Constitutively active STING causes neuroinflammation and degeneration of dopaminergic neurons in mice

  1. Eva M Szego
  2. Laura Malz
  3. Nadine Bernhardt
  4. Angela Rösen-Wolff
  5. Björn H Falkenburger  Is a corresponding author
  6. Hella Luksch
  1. Department of Neurology, TU Dresden, Germany
  2. Departments of Neurology & Pediatrics, TU Dresden, Germany
  3. Department of Psychiatry, TU Dresden, Germany
  4. Department of Pediatrics, TU Dresden, Germany
  5. Deutsches Zentrum für Neurodegenerative Erkrankungen, Germany
11 figures, 1 table and 2 additional files

Figures

Figure 1 with 1 supplement
Constitutive STING activation induces neuroinflammation and neurodegeneration in adult mice.

(A) Representative images of striatal sections from STING WT and STING ki mice stained for the microglia marker Iba1. Scale bar: 50 μm. (B) Representative images of striatal sections stained for the astroglia marker GFAP. Scale bar: 50 μm. (C) Representative images of midbrain sections containing the substantia nigra (SN) from STING WT and STING ki mice (stitched from two microscopy fields) stained for tyrosine hydroxylase (TH). Scale bar: 100 μm. (D) Representative images of striatal sections stained for TH from STING WT and STING ki mice. Scale bar: 10 μm. (E) Area fraction positive for Iba1, normalized to the mean of STING WT mice. Markers represent individual animals (black: STING WT animals, red: STING ki animals). Lines represent mean ± SD. Comparison by t-test (***: p=0.0007, n=5). Graph showing the counted numbers of Iba1-positive neurons is on Figure 1—figure supplement 1A. (F) Area fraction positive for GFAP, normalized to the mean of STING WT mice (**: p=0.0011; t-test, n=5). Graph showing the counted numbers of GFAP-positive neurons is on Figure 1—figure supplement 1B. (G) Number of TH-positive neurons (*: p=0.0257; t-test, n=5). Graph showing the counted numbers of dopaminergic neurons is on Figure 1—figure supplement 1D. (H) Area fraction positive for TH (**: p=0.0081; t-test, n=5). (I) Concentration of dopamine (*: p=0.0448; t-test, n=5) in striatal lysates from STING WT and STING ki animals, normalized to the mean concentration in STING WT. Graph showing quantification of the dopamine metabolites is in Figure 1—figure supplement 1E.

Figure 1—figure supplement 1
Neuroinflammation and neurodegeneration in adult mice.

(A) Number of Iba1-positive microglia in the striatum of adult mice (P=0,0000, t-test). (B) Number of GFAP-positive astroglia in the striatum of adult mice (p=0.0029, t-test). (C) Representative images on the ventral midbrain containing the substantia nigra of STING WT and STING ki mice, stained for TH (cyan), Iba1 (magenta) and GFAP (green). Scale bar: 100 µm. Insets show grayscale signal of Iba1 or GFAP staining. (D) Number of TH-positive neurons in the substantia nigra of adult mice (p=0.0257; t-test). (E) Dopamine metabolism (concentration of dopamine metabolites DOPAC +HVA) / dopamine in adult mice (p=0.0179; t-test). (F) Area fraction positive for GFAP signal in the substantia nigra as relative to the mean of STING WT (**: p=0.0078; t-test). (G) Area fraction positive for Iba1 signal in the substantia nigra as relative to the mean of STING WT (**: p=0.0078; t-test). (H) Number of counted GFAP-positive cells in the substantia nigra of adult mice (p=0.0003, t-test). (I) Number of counted Iba1-positive cells in the substantia nigra of adult mice (p=0.0000, t-test). n=5 for all graphs.

Figure 2 with 1 supplement
Neuroinflammation without neurodegeneration in juvenile mice with constitutive STING activation.

(A) Representative images of striatal sections stained for the microglia marker Iba1 from 5-week-old STING WT and STING ki mice. Scale bar: 50 μm (B) Representative images of striatal sections stained for the astroglia marker GFAP from 5-week-old STING WT and STING ki mice. Scale bar: 50 μm (C) Representative images of midbrain sections containing the substantia nigra (SN, stitched from two microscopy fields) stained for tyrosine hydroxylase (TH) from 5-week-old STING WT and STING ki mice. Scale bar: 100 μm (D) Representative images of striatal sections stained for TH from 5-week-old STING WT and STING ki mice. Scale bar: 10 μm. (E) Area fraction positive for Iba1, normalized to the mean of STING WT (***: p=0,0009; t-test, n=5–6). (F) Area fraction positive for GFAP, normalized to the mean of STING WT brains (***: p=0.0007; t-test, n=5–6). (G) Number of TH-positive neurons (mean ± SD; t-test). Graph showing the counted numbers of dopaminergic neurons is on Figure 2—figure supplement 1A. (H) Area fraction positive for TH (mean ± SD, t-test, n=5–6). (I) Dopamine concentration in striatal lysates from 5-week-old STING WT and STING ki mice, measured by HPLC and normalized to the mean of STING WT (mean ± SD, t-test, n=5–6). Dopamine metabolites are in Figure 2—figure supplement 1B.

Figure 2—figure supplement 1
Neuroinflammation without neurodegeneration in juvenile mice.

(A) Number of TH-positive neurons in the substantia nigra of juvenile mice (p=0.5188; t-test). (B) Dopamine metabolism in juvenile mice (p=0.9545; t-test). n=5–6.

Figure 3 with 1 supplement
Constitutive STING activation induces alpha-synuclein pathology and synapse loss in adult mice.

(A) Representative western blot images showing phosphorylated alpha-synuclein (S129; paSyn) and the loading control βIII-tubulin detected from the substantia nigra (upper panel) and striatum (lower panel). Total levels of aSyn detected from the same membranes are shown on Figure 3—figure supplement 1A, B. (B) Ratio of paSyn and total aSyn signals, expressed as relative to the mean of STING WT (substantia nigra (SN): p=0.000065; striatum: p=0.019; t-test, n=5). (C) Representative western blot images showing aSyn and βIII-tubulin detected from the Triton X-100 insoluble (upper panel) and soluble (lower panel) fractions prepared from the substantia nigra or from the striatum. (D) Ratio of aSyn signals detected in the Triton X-100 soluble and insoluble fractions, expressed as relative to the mean of STING ki (substantia nigra (SN): p=0.00001; striatum: p<0.00001; t-test, n=6). (E) Representative images of striatal sections from 20-week-old STING WT and STING ki mice stained with Thioflavin S (magenta) and nuclear sdye (blue) on the composite images, and ThioS BW. Scale bar: 20 μm. (F) Number of cells with inclusions positive for Thioflavin S (ThioS) per mm2 (*: p=0.0141; t-test, n=5). (G) Representative images of striatal sections from 20-week-old STING WT and STING ki mice stained for the presynaptic marker synapsin (upper panel) or for the post-synaptic marker homer (lower panel). Scale bar: 10 μm. (H–I) Area fraction positive for synapsin (H, p=0.0053) or homer (I, p=0.0408) (mean ± SD; t-test, n=5).

Figure 3—figure supplement 1
Uncropped membranes showing α-Synuclein signal detected from whole cell lysates of striatum and substantia nigra.

(A) Representative Western blot membrane showing aSyn signal in lysates prepared from the substantia nigra of STING WT and STING ki mice. (B) Representative Western blot membrane showing aSyn signal in lysates prepared from the striatum of STING WT and STING ki mice. (C) Ratio of aSyn and βIII tubulin in lysates prepared from the substantia nigra (SN) or striatum, expressed as relative to the mean of STING WT (SN: *: p=0.0462; STR: *: p=0.0491, t-test, n=5).

Figure 3—figure supplement 1—source data 1

Uncropped membranes showing α-Synuclein signal detected from whole cell lysates of striatum and substantia nigra.

https://cdn.elifesciences.org/articles/81943/elife-81943-fig3-figsupp1-data1-v2.zip
Figure 4 with 1 supplement
Activation of IFN and NF-κB/inflammasome related genes in the striatum and SN of STING ki mice.

(A–C) Expression of ISGs in the substantia nigra of STING WT and STING ki mice. (A) Ifi44 (p=0.1552, n=4–5), (B) Mx1 (Mann Whitney test, p=0.9048, n=4–5), (C) Sting1 (P=0.9184, n=4–5). (D–F) Expression of NF-κB/inflammasome related genes in the substantia nigra of STING WT and STING ki mice. (D) Tnfa (p=0.0691, n=4–5), (E) Il1b (Mann Whitney test, *: p=0.0159, n=4–5), (F) Casp1 (**: p=0.0018, n=4–5). (G–L) Expression of ISGs in the striatum of STING WT and STING ki mice. (G) Ifi44 (**: p=0.0078, n=3–4), (H) Mx1 (*: p=0.0183, n=3–4), (I) Sting1 (*: p=0.024, n=3–4). (J–L) Expression of NF-κB/inflammasome related genes in the striatum of STING WT and STING ki mice. (J) Tnfa (n=3–4), (K) Il1b (*: p=0.0397, n=3–4), (L) Casp1 (**: p=0.0017, n=3–5). Markers represent individual animals, bars represent mean ± SD. Analysis was t-test, if not indicated otherwise.

Figure 4—figure supplement 1
Expression of IFN and NF-κB and inflammasome dependent genes in the cortex of juvenile and adult STING ki mice.

(A–D) Expression of ISGs in the frontal cortex of STING WT and STING ki mice. (A) Ifi44 (***: p=0.0002277; *: p=0.044987, n=5), (B) Mx1 (***: p=0.0000003; *: p=0.016835; for comparison between age groups ***: p=0.000602, n=4–5), (C) Cxcl10 (***: p=0.000001; **: p=0.0017215; for comparison between age groups **: p=0.001483, n=4–5), (D) Sting1 (*: p=0.0184042, n=4–5). (E–G) Expression of NF-κB/inflammasome related genes in the frontal cortex of STING WT and STING ki mice. (E) Tnfa (***: p=0.0001448, for comparison between age groups *: p=0.03952, n=5). (F) Il1b (***: p=0.00005, *: p=0.0389, for comparison between age groups **: p=0.00241, n=4–5). (G) Casp1 (***: p=0.0000369, for comparison between age groups ***: p=0.000064, n=4–5). Markers represent individual animals, bars represent mean ± SD. Analysis was two-way ANOVA with Tukey HSD post-hoc test.

Figure 5 with 1 supplement
Nuclear translocation of pSTAT3 and NF-κB in the striatum of 5-week-old and 20 week-old STING WT and STING ki mice.

(A) Representative images of striatal sections from 5-week-old (upper images) and 20-week-old (lower images) STING WT and STING ki mice stained for Iba1 (green), GFAP (white) and phosphorylated-STAT3 (pSTAT3; red). Images show color coded merged channels (center) and in addition pSTAT3 staining in grayscale (left and right). Scale bar: 10 μm. (B) Representative images of striatal sections from 5-week-old (upper images) and 20 week-old (lower images) STING WT and STING ki mice stained for Iba1 (green), GFAP (white), and NF-κB (red). NF-kB staining is shown in grey in separate images. Scale bar: 10 μm. (C) Number of pSTAT3-positive nuclei/mm3 (***: p=0.00004; **: p=0.0025 for the interaction; two-way ANOVA, Bonferroni post-hoc test, n=5). (D) Number of NF-kB-positive nuclei/mm3 (**: p=0.009; ***: p=0.0007; mean ± SD; two-way ANOVA, Bonferroni post-hoc test, n=5).

Figure 5—figure supplement 1
Uncropped membranes showing the levels of inflammatory proteins in the striatum.

(A) Representative images of striatal sections from 20-week-old STING WT and STING ki mice stained for phosphorylated-STAT3 (pSTAT3, magenta), Iba1 (blue) and NeuN (green). Images show color-coded merged channels (left) and in addition all channels in grayscale (right). Scale bar: 20 μm. (B) Ratio of NeuN-positive cells positive for pSTAT3 as well: (number of pSTAT3- and NeuN-positive cells)/(total number of NeuN-positve cells); (****: p=0.000065; t-test). (C) Ratio of neuronal pSTAT3-positive nuclei: (number of pSTAT3- and NeuN-positive cells)/(total number of pSTAT3-positve cells); **: p=0.0025; t-test. n=4.

Figure 6 with 1 supplement
Activation of IRF3 and NF-kB-related signalling pathways in double transgenic mice with STING N153S/WT ki and knock-out for Ifnar1 or Caspase-1.

(A) Representative western blot images showing interferon regulatory factor 3 (IRF3, upper panel), the nuclear marker histone deacetylase 1 (HDAC1, middle panel) and cytoplasmic marker glycerinaldehyd-3-phosphat-dehydrogenase (GAPDH, lower panel) detected from the striatum. Images of the whole membrane stained for the different proteins are shown on Figure 6—figure supplement 1A-C. (B) Ratio of IRF3 and HDAC1, expressed as relative to the mean of STING WT (****: p<0.00001; **: p=0.0043; two-way ANOVA with Tukey post-hoc test, n=4). Ratio of IRF3 and GAPDH expressed as relative to the mean of STING WT is shown on Figure 6—figure supplement 1D. (C) Representative western blot images showing NLR family pyrin domain containing 3 (NLRP3, upper panel), Il1b and pro-Il1b (middle panel) and the loading control βIII tubulin (lower panel) detected from the striatum. Images of the whole membrane stained for the different proteins are shown on Figure 6—figure supplement 1E-H. (D) Ratio of NLRP3 and βIII tubulin, expressed as relative to the mean of STING WT (**: p<0.0057; **: p=0.0029; two-way ANOVA with Tukey post-hoc test, n=4). (E) Ratio of Il1b and pro-Il1b, expressed as relative to the mean of STING WT (****: p<0.00001; two-way ANOVA with Tukey post-hoc test, n=4). (F) Representative images of striatal sections stained for the astroglia marker GFAP (green), microglia marker Iba1 (magenta), apoptosis-associated speck-like protein (ASC, cyan) and Hoechst (blue) from STING WT or STING ki mice on a background of interferon a receptor knockout (Ifnar1-/-), caspase-1 knockout (Casp1-/-) or Ifnar1+/+, Casp1+/+ (Ctrl.). Scale bar: 20 μm. Magnified insets show Iba1 and ASC, cyan arrowheads indicate ASC specks. (G) Area fraction of ASC signal within microglia as relative to the mean of STING WT (****: p<0.00001; two-way ANOVA with Tukey post-hoc test, n=4–5). (H) Number of ASC-positive dots within microglia as relative to the mean of STING WT (****: p<0.00001; two-way ANOVA with Tukey post-hoc test, n=4–5). (I) Number of ASC-positive dots within astroglia as relative to the mean of STING WT (**: p=0.0083; ***: p=0.0006; two-way ANOVA with Tukey post-hoc test, n=4–5). Graph showing area fraction of ASC signal within astroglia is on Figure 6—figure supplement 1I.

Figure 6—figure supplement 1
Uncropped membranes showing the levels of inflammatory proteins in the striatum.

(A–C) Images of the whole membrane developed for the HDAC1 (A), GAPDH (B) and IRF3 signals. (D) Ratio of IRF3 and GAPDH signal (‘cytoplasmic IRF3’) as relative to the mean of STING WT (two-way ANOVA with Tukey HSD post-hoc test, n=4). (E–H) Images of the whole membrane developed for the NLRP3 and Il1b with short exposure time (E) or longer exposure time (F); and for βIII tubulin (H). Membrane was cut between the NLRP3 and tubulin signals. (I) Area fraction positive for ASC signal within GFAP-positive astroglia, as relative to the mean of STING WT (****: p<0.00001; *: p=0.038, two-way ANOVA with Tukey HSD post-hoc test, n=4–5). (J) Mean intensity of ASC signal in GFAP-positive or in Iba1-positive cells, expressed as relative to astroglial ASC signal in STING WT animals (***: p=0.0001; ****: p=0.000054; interaction: ****: p=0.000078, n=4–5).

Figure 7 with 2 supplements
Neuroinflammation in adult double transgenic mice with STING ki and knock-out for Ifnar1 or Caspase-1.

(A) Heatmap showing z-scores of different inflammatory markers in double transgenic mice with STING N153S/WT ki and knock-out for Ifnar1 or Caspase-1. Graphs with data for each mediator individually are on (Figure 7—figure supplement 2). (B) Representative images of striatal sections stained for the microglia marker Iba1. Sections were obtained from adult STING WT (upper images) or STING ki (lower images) mice on a background of interferon a receptor knockout (Ifnar1-/-), caspase-1 knockout (Casp1-/-) or Ifnar1+/+, Casp1+/+ (Ctrl.). Scale bar: 50 μm. (C) Representative images of striatal sections stained for the astroglia marker GFAP from STING WT (upper images) or STING ki (lower images) mice on a background of interferon a receptor knockout (Ifnar1-/-), caspase-1 knockout (Casp1-/-) or Ifnar1+/+, Casp1+/+ (Ctrl.). Scale bar: 50 μm. (D) Area fraction positive for Iba1, normalized to the mean of STING WT brains (differences in +/+ mice ***: p=0.0000001; for Ifnar1-/- ***: p=0.000003; for Casp1-/- ***: p=0.0029374; two-way ANOVA with Bonferroni post-hoc test, n=5–6). (E) Area fraction positive for GFAP, normalized to STING WT on Ctrl. Background (***: p=0.0000 for STING WT vs STING ki on Ctrl.; ***: p=0.0000 on Ifnar1-/-; background, ***: p=0.0006 on Casp1-/- background; two-way ANOVA with Bonferroni post-hoc test, n=5–6).

Figure 7—figure supplement 1
Expression of IFN- and NF-κB/inflammasome related genes in the cortex of double transgenic mice with STING N153S/WT ki and knock-out for Ifnar1 or Caspase-1.

Gene expression in the cortex (two-way ANOVA with Tukey HSD post-hoc test). (A) Ifi44 (*: p=0.01414; **: p=0.0037655; for interaction between Ctrl. and Ifnar1-/-, **: p=0.005101, n=4–5). (B) Mx1 (*: p=0.02823; **: p=0.00573, n=4–5). (C) Il1b (Ctrl. background *: p=0.04534; **: p=0.005405; Casp1-/- background *: p=0.0107096; for interaction between Ctrl. and Casp1-/-, *: p=0.01298, n=4–5). (D) Cxcl10 (Ctrl. background **: p=0.0030844; Ifnar1-/- background: p=0.025893; Casp1-/- background **: p=0.0041598, n=4–5). (E) Tnfa (all differences n.s., n=4–5). (F) Sting1 (all differences n.s., n=4–5).

Figure 7—figure supplement 2
Levels of inflammatory mediators and gene expression in the striatum of double transgenic mice with STING N153S/WT ki and knock-out for Ifnar1 or Caspase-1.

(A–M) Tissue levels of different ‘M1’ and ‘M2’ mediators in the striatum as measured by LegendPlex Immunoassay (ANOVA with Tukey HSD post-hoc test). (A) IFNα (*: p=0.0394; for interaction: *: p=0.0465); (B) IFNγ (*: p=0.0194); (C) Cxcl9 (*: p=0.0129); (D) Cxcl10 (*: p=0.0223); (E) CCL3 (***: p=0.0009); (F) CCL2 (*: p=0.0264); (G) TNFα; (H) IL-6; (I) CCL4; (J) Il4; (K) IL-10; (L) GM-CSF; (M) VEGF; (N–Q) Gene expression in the striatum (two-way ANOVA with Tukey HSD post-hoc test). (N) Nos2 (*: p=0.0214). (O) Ym-1 (***: p=0.0004). (P) Il4. (Q) Retnla, b or g (p=0.021). (R) Principal component analysis (PCA) of immune mediators shows segregation of STING ki animals on the Ifnar1+/+, Casp1+/+ (ki) and on the Ifnar1-/- (Ifnar1-/- ki) from all other groups. n=4–6.

Oxidative stress in the striatum of double transgenic mice with STING N153S/WT ki and knock-out for Ifnar1 or Caspase-1.

(A) Mitochondrial reactive oxygen species (ROS) of striatal lysates determined using MitoSOX and expressed relative to the mean of STING WT (**: p=0.0072; ****: p<0.00001; two-way ANOVA with Tukey post-hoc test, n=4). (B) Cytoplasmic ROS of striatal lysates determined using DCFH-DA and expressed relative to the mean of STING WT (****: p<0.00001; *: p=0,034; **: p=0.0069; two-way ANOVA with Tukey post-hoc test, n=4). (C) Nitrite levels as markers of nitric oxide activity measured in striatal lysates (****: p<0.00001; **: p=0.0052; two-way ANOVA with Tukey post-hoc test, n=4–5).

Figure 9 with 1 supplement
Degeneration of dopaminergic neurons in double transgenic mice with STING N153S/WT ki and knock-out for Ifnar1 or Caspase-1.

(A) Representative images of striatal sections stained for tyrosine hydroxylase (TH) from STING WT (upper images) or STING ki (lower images) mice on a background of interferon a receptor knockout (Ifnar1-/-), caspase-1 knockout (Casp1-/-) or Ifnar1+/+, Casp1+/+ (Ctrl.). Scale bar: 10 μm. (B) Area fraction positive for TH, normalized to STING WT on Ctrl. background (***: p=0.0000 for Ctrl. background; *: p=0.043 for Ifnar1-/-; **: p=0.0126845 for Casp1-/-; for interaction between Ctrl. background and Ifnar1-/-: p=0.00157; between Ctrl. and Casp1-/-: p=0.007326; two-way ANOVA with Bonferroni post-hoc test, n=5–6). (C) Concentration of dopamine in striatal lysates of STING WT and STING ki mice, normalized to STING WT on Ctrl. background. (***: p=0.0005; t-test, n=5–6). Dopamine metabolism is shown on Figure 9—figure supplement 1.

Figure 9—figure supplement 1
Dopamine metabolism in the striatum of double transgenic mice with STING N153S/WT ki and knock-out for Ifnar1 or Caspase-1.

Dopamine metabolism in the striatum (**: p=0.0073; two-way ANOVA, Tukey HSD post-hoc test, n=3–8).

Author response image 1
Example for correct genotyping of animals used in the experiments.

(a) PCR product by using primers specifically detecting the N153S variant of STING. (b) PCR product by using primers specifically detecting the WT (mouse) variant of STING. Mice with ID 66, 69, 70 and 71 are STING ki expressing both the mutant Sting1 and the WT Sting1. Mice with ID 67, 68, 72, 73 are STING WT. (c) PCR product by using primers specifically detecting the mouse Ifnar. (d) PCR product by using primers specifically detecting the lack of mouse Ifnar. All mice are Ifnar-/-.

Author response image 2

Tables

Appendix 1—key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Strain (Mus musculus)STING ki (STING
N153S knock-in mice)
Luksch et al., 2019C57BL/6 N background,
not the same mouse line
as Warner et al., 2017
Strain (Mus musculus)Ifnar1-/-Siedel et al., 2020backcrossed 20 times to
C57BL/6
Strain (Mus musculus)Casp1-/-Reinke et al., 2020C57BL/6 background
AntibodyAnti-Iba1, rabbit polyclonalFujifilm Wako ChemicalsCat# 019–197411:1000
AntibodyAnti-GFAP, chicken polyclonalAbcamCat# ab46741:2000
AntibodyAnti- NFκB p65 (D14E12), rabbit polyclonalCell Signaling TechnologyCat# 82421:500
AntibodyAnti- Tubulin βIII, rabbit polyclonalCovanceCat# PRB-435P1:5000
AntibodyAnti-phospho-a-synuclein, rabbit monoclonalAbcamCat# ab512531:1000
AntibodyAnti-a-synuclein, mouse monoclonalBD BioscienceCat# 6107871:2000
AntibodyAnti- TH, sheep, polyclonalPel-FreezeCat# P401011:2000
AntibodyAnti-synapsin, chicken monoclonalSynaptic SystemsCat# 1600021:1000
AntibodyAnti-homer, rabbit monoclonalSynaptic SystemsCat# 1060061:1000
AntibodyAnti-ASC, rabbit monoclonalAdipogenCat# AG-25B-0006-C1001:700
AntibodyAnti-IL1b, rabbit monoclonalCell Signaling TechnologyCat# 20221:300
AntibodyAnti- NLRP3, mouse monoclonalAdipogenCat# AG-20B-000600061:1000
AntibodyAnti- NeuN, chicken polyclonalSynaptic SystemsCat# 2660061:1500
AntibodyAnti- HDAC1, mouse, monoclonalThermo FisherCat# MA5-18071:1000
AntibodyAnti- IRF3, rabbit polyclonalSanta CruzCat# sc-90821:1000
AntibodyAnti- GAPDH, mouse, monoclonalMilliporeCat# MAB3741:5000
AntibodyHRP conjugated donkey anti-mouse polyclonalJackson ImmunoResearchCat# 715-035-1501:5000
AntibodyHRP conjugated donkey anti-rabbit polyclonalJackson ImmunoResearchCat# 711-035-1521:5000
AntibodyAlexa 555 conjugated donkey anti-rabbit polyclonalInvitrogenCat# A315721:2000
AntibodyAlexa 647 conjugated donkey anti-chicken polyclonalJackson ImmunoResearchCat# 703-605-1551:2000
AntibodyAlexa 488 conjugated donkey anti-sheep polyclonalInvitrogenCat# A110151:2000
AntibodyAlexa 488 conjugated donkey anti-rabbit polyclonalInvitrogenCat# A212061:2000
Sequence-based reagentIfi44 fw primerThis paperPCR primerAACTGACTGCTC
GCAATAATGT
Sequence-based reagentIfi44 rev primerThis paperPCR primerGTAACACAGCA
ATGCCTCTTGT
Sequence-based reagentMx1 fw primerThis paperPCR primerAACCCTGCTACCTTTCAA
Sequence-based reagentMx1 rev primerThis paperPCR primerAAGCATCGTTT
TCTCTATTTC
Sequence-based reagentSting1 fw primerThis paperPCR primerCTGCTGACATAT
ACCTCAGTTG
Sequence-based reagentSting1 rev primerThis paperPCR primerGAGCATGTTGT
TATGTAGCTG
Sequence-based reagentCxcl10 fw primerThis paperPCR primerCCAAGTGCTGC
CGTCATTTTC
Sequence-based reagentCxcl10 rev primerThis paperPCR primerGGCTCGCAGG
GATGATTTCAA
Sequence-based reagentTnfa fw primerThis paperPCR primerCCTGTAGCCC
ACGTCGTAG
Sequence-based reagentTnfa rev primerThis paperPCR primerGGGAGTAGACA
AGGTACAACCC
Sequence-based reagentCasp1 fw primerThis paperPCR primerGCTGCCTGCCC
AGAGCACAAG
Sequence-based reagentCasp1 rev primerThis paperPCR primerCTCTTCAGAGTCTCTTACTG
Sequence-based reagentIl1b fw primerThis paperPCR primerGAAATGCCACC
TTTTGACAGTG
Sequence-based reagentIl1b rev primerThis paperPCR primerTGGATGCTCTCA
TCAGGACAG
Sequence-based reagentHprt1 fw primerThis paperPCR primerTCAGTCAACGGG
GGACATAAA
Sequence-based reagentHprt1 rev primerThis paperPCR primerGGGGCTGTACT
GCTTAACCAG
Sequence-based reagentRpl13 fw primerThis paperPCR primerAGCCTACCAGAA
AGTTTGCTTAC
Sequence-based reagentRpl13 rev primerThis paperPCR primerGCTTCTTCTTCC
GATAGTGCATC
Sequence-based reagentEef2 fw primerThis paperPCR primerCCGACTCCC
TTGTGTGCAA
Sequence-based reagentEef2 rev primerThis paperPCR primerAGTTCAGGTCG
TTCTCAGAGAG
Sequence-based reagentYm1 fw primerThis paperPCR primerCATTCAGTCAG
TTATCAGATTCC
Sequence-based reagentYm1 rev primerThis paperPCR primerAGTGAGTAGCAGCCTTGG
Sequence-based reagentIl4 fw primerThis paperPCR primerAGATGGATGTGCC
AAACGTCCTCA
Sequence-based reagentIl4 rev primerThis paperPCR primerAATATGCGAAGCA
CCTTGGAAGCC
Sequence-based reagentNos2 fw primerThis paperPCR primerCTGCTGGTGGTGAC
AAGCACATTT
Sequence-based reagentNos2 rev primerThis paperPCR primerATGTCATGAGCAAA
GGCGCAGAAC
Sequence-based reagentRetnl1 fw primerThis paperPCR primerTGGAGAATAAGGTCAAGGAAC
Sequence-based reagentRetnlb rev primerThis paperPCR primerGTCAACGAGTAAGCACAGG
Commercial assay or kitPierce Micro BCA Protein-Assay-KitThermo Fisher Scientific IncCat# 23235
Commercial assay or kitWesternBright Chemilumineszenz
Substrat Sirius
Biozym Scientific GmbHCat# 541019
Commercial assay or kitM-MLV Reverse Transriptase KitPromegaCat# M1701
Commercial assay or kitSV Total RNA Isolation SystemPromegaCat# Z3101
Commercial assay or kitGoTaq qPCR Master MixPromegaCat# A6002
Chemical compound, drugPhosSTOP phosphatase inhibitorSigma-AldrichCat# 4906845001
Chemical compound, drugPrecision Plus Protein WesternC Protein StandardsBioRadCat# 1610376
Chemical compound, drugRNasin Ribonuclease InhibitorPromegaCat# N2511
Chemical compound, drugDeoxynucleotide Triphosphates (dNTPs)PromegaCat# U1205
Chemical compound, drugM-MLV 5 x Reaction BufferPromegaCat# M1701
Chemical compound, drugFluoromount-GSouthern BiotechCat# 0100–01
Chemical compound, drugNucSpot Live 650 Nuclear StainBiotiumCat# 40082
Chemical compound, drugThioflavin SSigma-AldrichCat# T1892
Chemical compound, drugMitosoxThermo FisherCat# M36008
Chemical compound, drugDi(Acetoxymethyl Ester)
(6-Carboxy-2',7'-
Dichlorodihydrofluorescein
Diacetate) (DCFH-DA)
Thermo FisherCat# C2938
Software, algorithmGraphPad Prism 5. 01 and 9.0.0GraphPad Software
Software, algorithmImageJ FijiWayne Rasband (NIH)
Software, algorithmQuantStudio5 qPCR Data
Analysis Software
Thermo Fisher Scientific Inc
OtherQuantStudio 5 Real-Time
PCR System
Thermo Fisher Scientific Inc
OtherZeiss Axio Observer Z1Carl Zeiss
OtherZeiss Spinning DiscCarl Zeiss
OtherInfinite 200 M Multimode-
Plate-Reader
Tecan Group

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  1. Eva M Szego
  2. Laura Malz
  3. Nadine Bernhardt
  4. Angela Rösen-Wolff
  5. Björn H Falkenburger
  6. Hella Luksch
(2022)
Constitutively active STING causes neuroinflammation and degeneration of dopaminergic neurons in mice
eLife 11:e81943.
https://doi.org/10.7554/eLife.81943