The regional distribution of resident immune cells shapes distinct immunological environments along the murine epididymis

  1. Christiane Pleuger  Is a corresponding author
  2. Dingding Ai
  3. Minea L Hoppe
  4. Laura T Winter
  5. Daniel Bohnert
  6. Dominik Karl
  7. Stefan Guenther
  8. Slava Epelman
  9. Crystal Kantores
  10. Monika Fijak
  11. Sarina Ravens
  12. Ralf Middendorff
  13. Johannes U Mayer
  14. Kate L Loveland
  15. Mark Hedger
  16. Sudhanshu Bhushan
  17. Andreas Meinhardt
  1. Institute of Anatomy and Cell Biology, Unit of Reproductive Biology, Justus-Liebig-University Giessen, Germany
  2. Hessian Center of Reproductive Medicine, Justus-Liebig-University of Giessen, Germany
  3. ECCPS Bioinformatics and Deep Sequencing Platform, Max Planck Institute for Heart and Lung Research, Germany
  4. Ted Rogers Center of Heart Research, Peter Munk Cardiac Centre, Toronto General Hospital Research Institute, University Health Network, Canada
  5. Institute of Immunology, Hannover Medical School, Germany
  6. Institute of Anatomy and Cell Biology, Unit of Signal Transduction, Justus-Liebig-University of Giessen, Germany
  7. Department of Dermatology and Allergology, Philipps-University of Marburg, Germany
  8. Centre of Reproductive Health, Hudson Institute of Medical Research, Australia
  9. Department of Molecular and Translational Sciences, School of Clinical Sciences, Monash Medical Centre, Monash University, Australia
7 figures, 1 table and 1 additional file

Figures

Figure 1 with 3 supplements
Analysis of differential immune responses of caput and cauda epididymides following uropathogenic Escherichia coli (UPEC) infection in C57BL/6J wild type mice.

(A) Male C57BL/6J mice (10–12 weeks of age) were intravasally injected with UPEC or saline vehicle alone (sham) after ligation of the vas deferens. For the study organs were harvested and analyzed …

Figure 1—figure supplement 1
Morphometric assessment of the differential immune responses within caput and cauda epididymides in C57BL/6J mice.

(A) Changes in weight (in mg) of caput and cauda epididymides throughout different infection time points post infection (mean ± SD, n=4–5, two-way ANOVA with Bonferroni post hoc test, *p<0.05, **p<0.…

Figure 1—figure supplement 2
Histological images (modified Masson-Goldner trichrome staining) of the epididymis of naïve, sham- and uropathogenic Escherichia coli (UPEC)-infected C57BL/6J mice at different time points (day 1, 5, 10 post infection).

Scale bar = 100 µm.

Figure 1—figure supplement 3
RNASeq analyses of sham- and uropathogenic Escherichia coli (UPEC)-infected C57BL/6J wild type mice.

(A) Principal component analysis of all investigated in vivo epididymitis samples in RNASeq analysis (n=3–4 per group). (B) Heatmap showing differentially expressed genes (DEG) between the caput of …

Figure 2 with 3 supplements
Analysis of changes in immune cell populations following infection with uropathogenic Escherichia coli (UPEC) in C57BL/6J wild type mice.

(A) Pearson’s correlation plot of infection time points (days post infection) and the area of immune cell infiltration within the total cauda area (%) determined by histological evaluation. Mean ± …

Figure 2—figure supplement 1
Gating strategy behind flow cytometry analyses of all immune cell populations under pathological conditions (displayed Figure 2).

Representative plots from a sham-infected corpus epididymis.

Figure 2—figure supplement 2
Infiltration of neutrophils in relation the bacterial appearance.

Percentage of infiltrating neutrophil granulocytes (blue) in relation to colony forming units (CFU)/mg tissue in proximal (IS/caput) and distal epididymis (corpus/cauda) of sham- and uropathogenic Es…

Figure 2—figure supplement 3
Multiplex assay-based determination of cytokine levels from ex vivo organ culture.

Related to Figure 2. (A) Indicated cytokines (IL-1α, IL-1β, TNFα, MCP-1, IL-6, IL-10) were measured within the culture media after ex vivo stimulation of the indicated epididymal regions with …

Figure 3 with 3 supplements
Single-cell RNA sequencing (scRNASeq) of different epididymal regions reveals immune cell heterogeneity within the murine epididymis under physiological conditions.

(A) Schematic overview of the experimental procedure for isolating extravascular CD45+ cells from different epididymal regions. (B) Uniform manifold approximation and projection (UMAP) plot of …

Figure 3—figure supplement 1
Extravascular CD45+ cells of different epididymal regions were sorted following the indicated gating strategy prior to single-cell RNASeq.
Figure 3—figure supplement 2
Quality controls for single-cell reads and re-confirmation of identified CD45+.

(A) Number of total cells, total genes and average reads per cells are indicated for different epididymal regions that were separately isolated. (B) Uniform manifold approximation and projection …

Figure 3—figure supplement 3
Expression of key marker genes for the identified immune cell populations within epididymal regions.

Uniform manifold approximation and projection (UMAP) plots showing the expression of selected key marker for the indicated immune cell population within the epididymal regions: initial segment, …

Figure 4 with 1 supplement
Quantification and localization of identified immune cell populations among epididymal regions (scale bar 20 µm).

(A) Distribution (assessed by flow cytometry n=4-8, bar diagram showing mean ± SD) of total leukocytes (CD45+ cells) and localization within the initial segment and corpus, as shown by …

Figure 4—figure supplement 1
Gating strategy behind flow cytometry analyses of all immune cell populations under physiological conditions.

(A) General gating that has been applied to each sample included exclusion of debris und sperm based on SSC-A vs. FSC-A, followed by a two-step single-cell gating (based on FSC and SSC), live cells …

Figure 5 with 2 supplements
Distinct macrophage subgroups exist within the murine epididymis.

(A) Uniform manifold approximation and projection (UMAP) plot and violin plots showing segregation of macrophages (clusters 1, 2, 7) and monocytes (clusters 8, 10) based on clustering and expression …

Figure 5—figure supplement 1
Uniform manifold approximation and projection (UMAP) plots showing the expression of selected key markers for identified macrophage subgroups, related to violin plots in Figure 4D.
Figure 5—figure supplement 2
Violin plots showing the expression of immediate-early activation genes (Fos, Jun, Egr1) as well as upregulated cytokines Tnf, Cxcl2, Ccl4 among identified macrophage subgroups.
Figure 6 with 4 supplements
Distribution and localization of identified macrophage subgroups by flow cytometry and immunofluorescence.

(A) Bar diagram showing the percentage of F4/80+ cells within the CD45+ population throughout the epididymal regions, assessed by flow cytometry (n=8, mean ± SD). (B) Stacked bar diagram displaying …

Figure 6—figure supplement 1
Gating strategy of macrophage subsets according to obtained single-cell RNA sequencing (scRNASeq) data.

Arrows indicate the gating strategy and identified subsets.

Figure 6—figure supplement 2
Macrophage subpopulations within CD45+ population.

Bar diagrams showing the percentage of identified macrophage subgroups within the CD45+ population throughout the epididymal regions, assessed by flow cytometry and mirroring the distribution …

Figure 6—figure supplement 3
Single channel reads of anti-F4/80 (purple) staining on epididymal cryo-sections from adult Cx3cr1GFPCcr2RFP reporter mice.

The majority of CX3CR1+ cells were F4/80+. Arrowheads indicate the small fraction of intraepithelial F4/80- CX3CR1+ cells within the initial segment (IS). Arrows indicate interstitial F4/80+ CX3CR1-

Figure 6—figure supplement 4
Single channel reads of anti-MHC-II (purple) staining on epididymal cryo-sections from adult Cx3cr1GFPCcr2RFP reporter mice.

Asterisks (*) indicate intraepithelial CX3CR1+MHC-II- cells within the initial segment (IS) and caput epididymides. Arrowheads indicate CX3CR1+MHC-II+ cells, lining the epididymal duct within the IS …

Figure 7 with 2 supplements
Resident macrophages differentially depend on monocytes within epididymal regions.

(A) Parabiosis was conducted by surgically conjoining wild type CD45.1+ donor mice with CD45.2 recipient Ccr2-/- mice for 6 months. Donor chimerism was confirmed on CD115+CD11b+Ly6Chi monocytes. (B) …

Figure 7—figure supplement 1
Gating strategy that was applied on blood samples from recipient Ccr2-/- mice.

General gating was initially performed based on FSC and SSC in order to exclude debris and doublets, before neutrophils were excluded by selecting Ly6G- cells. Monocytes were further gated using …

Figure 7—figure supplement 2
Representative plots (cauda) that were applied for the epididymal regions starting with general gating based on FSC and SSC in order to exclude debris and doublets.

Total CD64+CD11b+ cells were gated and further segregated using TIMD4 and CCR2. The ratio of CD45.1 and CD45.2+ events was assessed in CCR2+, TIMD4+, and CCR2-TIMD4- cells.

Tables

Appendix 1—key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Strain, strain background (Escherichia coli)CFT073Welch et al., 2002NCBI: txid19.9310Provided by T.Chakraborty, Justus-Liebig-University, Giessen, Germany
Strain, strain background (Mus musculus, male)C57BL/6 J wild typeCharles RiverJAX ID: 00066410–12 weeks old
Strain, strain background (Mus musculus, male)B6.129(Cg)-Cx3cr1tm1Litt
Ccr2tm2.1Ifc
/JernJ (Cx3cr1GFPCcr2RFP)
Jackson Laboratory Jung et al., 2000; Saederup et al., 2010JAX ID: 032127
Strain, strain background (Mus musculus, male)B6.SJL-Ptprca Pepcb/
BoyJ (B6 CD45.1)
Jackson Laboratory
(Janowska-Wieczorek et al., 2001;
Schluns et al., 2002;
Yang et al., 2002)
JAX ID: 002014
Strain, strain background (Mus musculus, male)B6.129S4-Ccr2tm1Ifc/J (Ccr2-/-)Jackson Laboratory
(Boring et al., 1997)
JAX ID: 004999
Strain, strain background (Mus musculus, male)C57/BL/6-Trdctm1Mal/J
(Trcd-H2BeGFP)
Jackson Laboratory
(Prinz et al., 2006)
JAX ID: 016941
AntibodyPerCP/C5.5 anti-mouse
CD45 (rat monoclonal)
BioLegendCat.No.: 103131;
RRID: AB_893344
FC (1:200) for infection study
AntibodyAF700 anti-mouse CD3 (rat monoclonal)BioLegendCat.No.: 100216;
RRID: AB_493697
FC (1:50) for infection study
AntibodyBrilliant Violet 511 anti-mouse/ human CD45R/B220 (rat monoclonal)BioLegendCat.No.: 103247;
RRID: AB_2561394
FC (1:200) for infection study
AntibodyBrilliant Violet 605 anti-
mouse NK1.1 (mouse
monoclonal)
BioLegendCat. No.: 108753;
RRID: AB_2686977
FC (1:200) for infection study
AntibodyBrilliant Violet 650 CD11c
(Armenian Hamster
monoclonal)
BioLegendCat.No.: 117339;
RRID: AB_2562414
FC (1:100) for infection study
AntibodyBrilliant Violet 711 anti-mouse
Ly-6G/Ly-6C (GR-1) (rat monoclonal)
BioLegendCat.No.: 108443;
RRID: AB_2562549
FC (1:200) for infection study
AntibodyBrilliant Violet 785 anti-mouse
I-A/I-E (MHC-II) (rat monoclonal)
BioLegendCat.No.: 107645;
RRID: AB_2565977
FC (1:200) for infection study
AntibodyPE/Dazzle594 anti-mouse
F4/80 (rat monoclonal)
BioLegendCat.No.:123145;
RRID: AB_2564132
FC (1:100) for infection study
AntibodyPE/Cyanine7 anti-mouse
CX3CR1 (mouse monoclonal)
BioLegendCat.No.:149015;
RRID: AB_2565699
FC (1:1000) for infection study
AntibodyFITC anti-mouse CCR2
(rat monoclonal)
BioLegendCat. No.: 150608;
RRID: AB_2616980
FC (1:200) for infection and steady-state study
AntibodyBrilliant Violet 421 anti-mouse
CX3CR1 (mouse monoclonal)
BioLegendCat.No.: 149023;
RRID: AB_2565706
FC 1:1000 for steady-state study
AntibodyPerCP/Cyanine5.5 anti-mouse
CD45 (rat monoclonal)
BioLegendCat.No.:157207;
RRID: AB_2860727
FC 1:100 for steady-state study
AntibodyPE/Cyanine7 anti-mouse
F4/80 (rat monoclonal)
BioLegendCat.No.: 123113;
RRID: AB_893490
FC 1:100 for steady-state study
AntibodyAPC anti-mouse CD163
(rat monoclonal)
BioLegendCat.No.:155305;
RRID: AB_2814059
FC 1:200 for steady-state study
AntibodyAPC/Cy7 anti-mouse I-A/I-E
(rat monoclonal)
BioLegendCat.No.: 107627;
RRID: AB_1659252
FC 1:200 for steady-state study
AntibodyBrilliant Violet 421 anti-mouse
NK1.1 (mouse monoclonal)
BioLegendCat.No.: 108731;
RRID: AB_10895916
FC 1:200 for steady-state study
AntibodyFITC anti-mouse CD3
(rat monoclonal)
BioLegendCat.No.: 100203;
RRID: AB_312660
FC 1:50 for steady-state study
AntibodyPE anti-mouse B220
(CD45R) (rat monoclonal)
MiltenyiCat.No.: 130-120-077;
RRID: AB_2751992
1:50 for steady-state study
AntibodyPE/Cyanine7 anti-mouse
TCRbeta chain (Armenian
Hamster monoclonal)
BioLegendCat.No. 109221;
RRID: AB_893627
1:100 for steady-state study
AntibodyAPC anti-mouse TCR g/d
(Armenian Hamster monoclonal)
BioLegendCat.No.: 118115;
RRID: AB_1731824
1:100 for steady-state study
AntibodyAPC/Fire750 anti-mouse
CD45 (mouse monoclonal)
BioLegendCat.No.: 103153;
RRID: AB_2572115
1:100 for steady-state study
AntibodyBrilliant Violet 421 anti-
mouse F4/80 (mouse monoclonal)
BioLegendCat.No.: 123131;
RRID: AB_10901171
1:100 for steady-state study
AntibodyPE anti-mouse CD209a
(DC-Sign) antibody
(mouse monoclonal)
BioLegendCat.No.: 833003;
RRID: AB_2721636
FC 1:50 for steady-state study
AntibodyPerCP/Cy5.5 anti-mouse/
human CD11b (rat monoclonal)
BioLegendCat.No.: 101228;
RRID: AB_893232
FC 1:200 for steady-state study
AntibodyAPC anti-mouse I-Ab
(mouse monoclonal)
BioLegendCat.No:116418;
RRID: AB_10574160
FC 1:200 for steady-state study and parabiosis
AntibodyPerCP/Cyanine5.5 anti-
mouse CD45.1 (mouse monoclonal)
BioLegendCat.No.: 110728;
RRID: AB_893346
FC 1:100 for steady-state study and parabiosis
AntibodyAPC/Cyanine7 anti-mouse
CD45.2 (mouse monoclonal)
BioLegendCat.No.: 109824;
RRID: AB_830789
i.v. injection 1:100
AntibodyPE anti-mouse CD64
(FcγRI) (mouse monoclonal)
BioLegendCat.No:. 139303;
RRID: AB_10612740
FC 1:100 for steady-state study and parabiosis
AntibodyPE/Cyanine7 anti-mouse
Tim-4 (rat monoclonal)
BioLegendCat.No.: 130010;
RRID: AB_2565719
FC 1:100 for parabiosis
AntibodyBrilliant Violet 785 anti-
mouse/human CD11b (rat monoclonal)
BioLegendCat.No.: 101224;
RRID: AB_755986
FC 1:100 for parabiosis
AntibodyAlexa Fluor 700 anti-mouse Ly-6G (rat monoclonal)BioLegendCat.No.: 127622;
RRID: AB_10643269
FC 1:100 for parabiosis
AntibodyPE anti-mouse CD115 (CSF-1R) (rat monoclonal)BioLegendCat.No.: 135506;
RRID: AB_1937253
FC 1:100 for parabiosis
AntibodyFITC anti-mouse Ly-6C (rat monoclonal)BioLegendCat.No.: 128006;
RRID: AB_1186135
FC 1:100 for parabiosis
Antibodyanti-mouse F4/80 (rat monoclonal)BioLegendCat.No. MCA497G;
RRID: AB_872005
IF 1:200
Antibodyanti-mouse Ly-6G+Ly-6C (rat monoclonal)abcamCat.No. ab25377;
RRID: AB_470492
IF 1:500
AntibodyPurified anti-mouse I-A/I-E
(mouse monoclonal)
BioLegendCat.No.: 107601;
RRID: AB_313316
IF 1:200
AntibodyPurified anti-mouse Ly-6C
(rat monoclonal)
BioLegendCat.No. 128002
RRID: AB_1134214
IF 1:100
AntibodyPurified anti-mouse CD3
(mouse monoclonal)
BioLegendCat.No. 100202
RRID: AB_312659
IF 1:50
AntibodyAnti-mouse Clec9a (sheep polyclonal)R&D SystemsCat.No. AF6776
RRID: AB_10890771
IF 1:50
Antibodyanti-mouse CD163 [TNKUPJ]
(rat monoclonal)
Invitrogen/ eBioscienceCat.No. 14-1631-82
RRID: AB_2716934
IF 1:200
Antibodyanti-NCR1 antibody
[EPR23097-35] (rabbit monoclonal)
abcamCat.No. ab233558
RRID: AB_2904203
IF 1:50
AntibodyAni-mouse DC-Sign (DC28)
(mouse monoclonal)
Santa CruzCat.No. sc-65740
RRID: AB_1121347
IF 1:50
AntibodyGoat anti-rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488InvitrogenCat-No. 11008
RRID: AB_143165
IF 1:2000
AntibodyGoat anti-rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 546InvitrogenCat.No. A-11010
RRID: AB_2534077
IF 1:2000
AntibodyGoat anti-rat IgG (H+L) Cross-Adsorbed secondary Antibody, Alexa Fluor 546InvitrogenCat.No. A-11081
RRID: AB_141738
IF 1:2000
AntibodyGoat Anti-Rat IgG H+L Alexa Fluor 647abcamCat.No. ab150159;
RRID: AB_2566823
IF 1:2000
AntibodyDonkey Anti-Sheep IgG (H+L) Alexa fluor 546InvitrogenCat.No.:A-21098
RRID: AB_2535752
IF: 1:2000
AntibodyBrilliant Violet 421 Mouse
IgG2a, κ Isotype Ctrl antibody
BioLegendCat.No.: 400259;
RRID: AB_10895919
FC 1:200 Isotype control
AntibodyBrilliant Violet 421 Rat IgG2a, κ Isotype Ctrl antibodyBioLegendCat.No.: 400535;
RRID: AB_10933427
FC 1:200 Isotype control
AntibodyBrilliant Violet 510 Rat IgG2a, κ, Isotype Ctrl antibodyBD BioscienceCat.No. 562952;
RRID: AB_2869438
FC 1:200 Isotype control
AntibodyFITC Rat IgG2b, κ Isotype Ctrl antibodyBioLegendCat.No.: 400605;
RRID: AB_326549
FC 1:200 Isotype control
AntibodyAlexa Fluor 488 Rat IgG2a, κ Isotype Ctrl antibodyBioLegendCat.No.: 400525;
RRID: AB_2864283
FC 1:200 Isotype control
AntibodyAlexa Fluor 488 Rat IgG2b, κ Isotype Ctrl antibodyBioLegendCat.No.: 400625;
RRID: AB_389321
FC 1:200 Isotype control
AntibodyPE Isotype Control Antibody, Rat IgG2aMiltenyiCat.No.: 130-123-747;
RRID: AB_2857628
FC 1:200 Isotype control
AntibodyPE Rat IgG2a, κ Isotype Ctrl antibodyBioLegendCat.No.: 400507;
RRID: AB_326530
FC 1:200 Isotype control
AntibodyPE Mouse IgG2a, κ Isotype Ctrl antibodyBioLegendCat.No.: 400213;
RRID: AB_2800438
FC 1:200 Isotype control
AntibodyPE Rat IgG2b kappa Isotype ControleBioscienceCat.No.: 12-4031-82;
RRID: AB_470042
FC 1:200 Isotype control
AntibodyPerCP/Cyanine5.5, Rat IgG2b, κ Isotype Ctrl antibodyBioLegendCat.No. 400631;
RRID: AB_893693
FC 1:200 Isotype control
AntibodyPE/Cyanine 7 Mouse IgG1, κ Isotype Ctrl antibodyBioLegendCat.No.: 400125;
RRID: AB_2861533
FC 1:200 Isotype control
AntibodyPE/Cyanine 7 Rat IgG2a, κ Isotype Ctrl antibodyBioLegendCat.No.: 400521; RRID: AB_326542FC 1:200 Isotype control
AntibodyPE/Cyanine 7 Armenian Hamster IgG Isotype Ctrl antibodyBioLegendCat.No.:400921; RRID: AB_2905473FC 1:200 Isotype control
AntibodyAPC Rat IgG2a, κ Isotype Ctrl antibodyBioLegendCat.No.: 400511; RRID: AB_2814702FC 1:200 Isotype control
AntibodyAPC Armenian Hamster IgG Isotype Ctrl antibodyBioLegendCat.No.: 400911; RRID: AB_2905474FC 1:200 Isotype control
AntibodyAPC Mouse IgG2a,
κ Isotype Ctrl (FC) antibody
BioLegendCat.No.: 400221; RRID: AB_2891178FC 1:200 Isotype control
AntibodyAPC/Cyanine7 Rat IgG2b,
κ Isotype Ctrl antibody
BioLegendCat.No.: 400628; RRID: AB_326565FC 1:200 Isotype control
AntibodyAPC/Fire750 Rat IgG2b,
κ Isotype Ctrl antibody
BioLegendCat.No.: 400669; RRID: AB_2905475FC 1:200 Isotype control
Commercial assay or kitM.O.M. (Mouse on Mouse)
Immunodetection Kit
Vector LaboratoriesCat.No. BMK-2202
Commercial assay or kitRNeasy Mini KitQiagenCat.No.: 74004
Commercial assay or kitLEGENDPlex with Mouse Inflammation PanelBioLegendCat.No.: 740446
Commercial assay or kitSMARTer Stranded Total
RNA-Seq Kit – Pico Input
Mammalian
TakaraCat.No.: 634488
Chemical compound, drugCollagenase D from Clostridium histolyticumRocheCat.No. 11088858001
Chemical compound, drugDNase ISigmaCat.No. D4513
Chemical compound, drugHyaluronidase type I-SSigmaCat.No. H3506
Chemical compound, drugRBC Lysis SolutionQiagenCat.No.: 158904
Chemical compound, drugUltraPure Lipopolysaccharide
from Escherichia coli O55:B5
SigmaCat.No.: L2880
Chemical compound, drugGibco RPMI1640 mediaFisher ScientificCat-No.: 11530586
Chemical compound, drugQIAzol Lysis ReagentQiagenCat.No.: 79306
Chemical compound, drugFc blocking reagentMiltenyiCat.No. 130-092-575
Chemical compound, drugGentamicin solutionSigma/ MerckCat.No. G1397
Other (dyes)ZombieAquaBioLegendCat.No.: 423101
Other (dyes)ZombieNIRBioLegendCat. No.: 423105
Other (dyes)Viobility 405/452 Fixable DyeMiltenyiCat.No.: 130-092-575
Other (dyes)DAPIInvitrogenD13061 µg/ml
OtherProLong Antifade Gold with DAPIInvitrogenP36931
OtherProLong Antifade Gold w/o DAPIInvitrogenP36930
Software, algorithmFlowJo v10.8.2BD Life SciencesRRID: SCR_008520https://www.flowjo.com/
Software, algorithmAdobe Illustrator 2020 (v24.0.1)AdobeRRID: SCR_010279https://www.adobe.com/de/products/illustrator.html
Software, algorithmGraphPad Prism v5GraphPad SoftwareRRID: SCR_002798https://www.graphpad.com/scientific-software/prism/
Software, algorithmInkScape v0.92.4The Inkscape ProjectRRID: SCR_014479https://inkscape.org/de/release/inkscape-0.92.4/
Software, algorithmImageJ 1.53 aWayne Rasband National Institute of Health, USARRID: SCR_003070https://imagej.nih.gov/ij/index.html
Software, algorithmZen 2.3 Version 14.0.26.201Carl Zeiss Microscopyhttps://www.zeiss.de/mikroskopie/produkte/mikroskopsoftware/zen-lite/zen-lite-download.html
Software, algorithmLEGENDPlex Software v8.0BioLegendsoftware provied by
BioLegend as part of the
LegendPlex kit for protein analysis
https://www.biolegend.com/en-us/legendplex
Software, algorithmFastQCAndrews, 2010RRID: SCR_014583http://www.bioinformatics.babraham.ac.uk/projects/fastqc/
Software, algorithmSTAR 2.6.1dDobin et al., 2013RRID: SCR_004463https://github.com/alexdobin/STAR/releases?page=2
Software, algorithmSubread packageLiao et al., 2013RRID: SCR_009803http://subread.sourceforge.net/
Software, algorithmDESeq2 V1.18.1Love et al., 2014RRID: SCR_015687https://bioconductor.org/packages/release/bioc/html/DESeq2.html
OtherBD Aria FusionBD BioScienceN/AInstrument
OtherHomogenizer MM400RetschN/AInstrument
OtherLabChip Gx Touch 24Perkin ElmerN/AInstrument
OtherLeica Cryotome CM1850LeicaN/AInstrument
OtherLeica Microtome RM2255LeicaN/AInstrument
OtherMACS Quant Analyzer 10MiltenyiN/AInstrument
OtherBD LSRFortessa Cell AnalyzerBD BiosciencesN/AInstrument
OtherNextSeq500IlluminaN/AInstrument
OtherZeiss LSM710 Confocal
Microscope
Carl Zeiss MicroscopyN/AInstrument
OtherOlympus BX51OlympusN/Ainstrument

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