Nuclear NAD+-biosynthetic enzyme NMNAT1 facilitates development and early survival of retinal neurons
- Department of Ophthalmology and Visual Sciences, Eye Institute, One Medical Center Drive, West Virginia University, United States
- Department of Biochemistry, One Medical Center Drive, West Virginia University, United States
Figures
Figure 1 with 1 supplement
Loss of NMNAT1 leads to early and severe retinal degeneration.
(A) Schematic depicting retina-specific Six3-Cre mediated excision of a segment of the Nmnat1 gene. (B) Relative Nmnat1 expression in retina from P4 knockout (-/-) and littermate control (+/+) mice as assessed by RT-qPCR (grey bars represent mean, significance determined using Mann-Whitney U test, n = 3 biological replicates). (C) Representative western blot showing levels of NMNAT1 and β-tubulin loading control in retinal lysate from P0 knockout and control mice. (D–G) Representative H&E-stained retinal cross sections from knockout and control mice at indicated ages. (H–J) Spider plots depicting mean retinal thickness at P0, P4, and P10. Data are represented as mean ± SD. *p < 0.05 using Student’s t-test, n = 3 biological replicates per age. Scale bars, 30 μm. Abbreviations: LP, loxP site; E1-4, exon 1–4; P, postnatal day; GCL, ganglion cell layer; NBL, neuroblastic layer; IPL, inner plexiform layer; OPL, outer plexiform layer; INL, inner nuclear layer; ONL, outer nuclear layer; IS/OS, photoreceptor inner segment/outer segment layer.
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Figure 1—source data 1
Quantification of Nmnat1 mRNA levels in P0 WT and KO retinas.
Numerical source data for retinal thickness quantification in P0, P4, and P10 WT and KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Validation of anti-NMNAT1 antibody in cell lines and retinal tissue.
(A) western blot of lysate of HEK293T cells transiently transfected with FLAG-tagged NMNAT1 demonstrating specificity of NMNAT1 antibody. (B) Western blot from Figure 1C, with dotted line indicating crop area. Arrowheads denote bands corresponding to NMNAT1.
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Figure 1—figure supplement 1—source data 1
Raw western blot scan of P4 WT and KO retinal lysate stained with anti-NMNAT1 antibody (700 channel).
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig1-figsupp1-data1-v2.zip
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Figure 1—figure supplement 1—source data 2
Raw western blot scan of P4 WT and KO retinal lysate stained with anti-beta-tubulin antibody (800 channel).
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig1-figsupp1-data2-v2.zip
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Figure 1—figure supplement 1—source data 3
Raw western blot scan of HEK293T cell lysate stained with anti-NMNAT1 antibody (700 channel).
Band corresponding to NMNAT1 is clearly labeled.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig1-figsupp1-data3-v2.zip
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Figure 1—figure supplement 1—source data 4
Raw western blot scan of HEK293T cell lysate stained with anti-FLAG antibody (800 channel).
Band corresponding to FLAG is clearly labeled.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig1-figsupp1-data4-v2.zip
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Figure 1—figure supplement 1—source data 5
Uncropped western blot scan of P4 WT and KO retinal lysate stained with anti-NMNAT1 antibody.
Band corresponding to NMNAT1 is clearly labeled.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig1-figsupp1-data5-v2.zip
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Figure 1—figure supplement 1—source data 6
Uncropped western blot scan of P4 WT and KO retinal lysate stained with anti-beta-tubulin antibody.
Band corresponding to beta-tubulin is clearly labeled.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig1-figsupp1-data6-v2.zip
Figure 2 with 1 supplement
NMNAT1 loss affects retinal bipolar, horizontal and amacrine cells.
Representative retinal sections from knockout (-/-) and floxed littermate control (+/+) mice at the indicated ages labeled with antibodies against BRN3A (A-D, green), Calbindin (CALB) (A–B, magenta) Calretinin (CALR) (E–H), and CHX10 (I–L). Quantification of BRN3A (M), CALB, (N), CALR (O), and CHX10-positive cells (P) are shown. In (O), only CALR-positive cells on the outer side of the IPL (layer indicated by white arrowheads) were counted. Data is represented as mean ± SD. n = 3 biological replicates for all panels; significance determined using Student’s t-test. Scale bars, 30 μm.
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Figure 2—source data 1
Numerical source data for retinal cell type quantification in P4 and P10 KO and WT retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Six3-Cre does not cause obvious defects in the mature retina.
Representative retinal sections from P65 wild-type (Nmnat1wt/wt) and Six3-Cre expressing (Nmnat1wt/wt;Six3-Cre) mice labeled with antibodies against calretinin (CALR) and BRN3A (A–B), recoverin (RCVRN) and rhodopsin (RHO) (C–D), synaptophysin (SYPH) (E,F), or CHX10 (G,H). Corresponding zoom panels are indicated with dotted rectangles. n = 3 biological replicates for all panels. Scale bars, 30 μm. Abbreviations: P, postnatal day; GCL, ganglion cell layer; IPL, inner plexiform layer; OPL, outer plexiform layer; INL, inner nuclear layer; ONL, outer nuclear layer; IS/OS, photoreceptor inner segment/outer segment layer.
Figure 3 with 5 supplements
NMNAT1 loss impairs photoreceptor terminal differentiation.
Representative retinal sections from knockout (-/-) and floxed littermate control (+/+) mice at the indicated ages labelled with antibodies against OTX2 (A–D), Phosducin (PDC) and peanut agglutinin (PNA) (E, F), M-opsin (OPN1MW) and PNA (G, H), recoverin (RCVRN) (H,I) or rhodopsin (RHO) (J, K). (L) Comparison of differentially expressed genes in E18.5 and P4 knockout retinas as assessed by RNA-sequencing. (M) GSEA of differentially expressed genes identified in E18.5 and P4 knockout retinas. (N, O) Relative expression of indicated genes in P4 and E18.5 knockout retinas as assessed by RNA-sequencing. n = 3 biological replicates for all panels, except (N), where n = 5 biological replicates. Corresponding zoom panels are indicated with dotted rectangles. Scale bars, 30 μm.
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Figure 3—source data 1
Gene ontology (GO) overrepresentation analysis of genes differentially expressed in KO retinas at E18.5 and P4 timepoints.
Numerical source data for expression (relative to WT levels) of select photoreceptor-specific genes in E18.5 and P4 KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Proliferation is only modestly affected in P0 NMNAT1 knockout retinas.
(A–B) Representative retinal sections from P0 knockout (-/-) and control (+/+) mice labelled with an antibody against phospho-histone H3 (PHH3). (C) Quantification of PHH3-positive nuclei. (D–E) Representative retinal sections from P4 knockout (-/-) and control (+/+) mice labelled with anti-mouse 488 secondary antibody. Representative retinal sections from P10 knockout and control mice labelled with antibodies against recoverin (RCVRN) (F,G) and rhodopsin (RHO) (H,I) are also shown. (J) RT-qPCR validation of indicated genes in P4 RNA-sequencing dataset. Corresponding zoom panels are indicated with dotted rectangles. Scale bars, 30 μm. Data is represented as mean ± SD. significance determined using Student’s t-test. n = 3 biological replicates for all panels (n = 5 biological replicates for RNA-seq data in (J)).
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Figure 3—figure supplement 1—source data 1
Numerical source data for expression (relative to WT) of select genes in P4 KO retinas using RNA-seq and RT-qPCR.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig3-figsupp1-data1-v2.xlsx
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Figure 3—figure supplement 1—source data 2
Numerical source data for quantification of PHH3-positive nuclei in P0 WT and KO retinal sections.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig3-figsupp1-data2-v2.xlsx
Figure 3—figure supplement 2
Global transcriptional changes in P4 NMNAT1 knockout retinas.
(A) Schematic depicting RNA-sequencing experimental approach for P4 knockout and control retinas. (B) Donut chart representing relative proportions of differentially upregulated and downregulated genes in P4 knockout retinas. (C) Results of pathway overrepresentation analysis of differentially expressed genes in P4 knockout retinas. n = 5 biological replicates.
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Figure 3—figure supplement 2—source data 1
Gene ontology (GO) overrepresentation analysis of significantly changed genes in P4 KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig3-figsupp2-data1-v2.xlsx
Figure 3—figure supplement 3
Global transcriptional changes in E18.5 NMNAT1 knockout retinas.
(A) Schematic depicting RNA-sequencing experimental approach for E18.5 knockout and control retinas. (B) Donut chart representing relative proportions of differentially upregulated and downregulated genes in E18.5 knockout retinas. (C) Results of pathway overrepresentation analysis of differentially expressed genes in E18.5 knockout retinas. n = 4 biological replicates (one outlier removed from knockout group).
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Figure 3—figure supplement 3—source data 1
Gene ontology (GO) overrepresentation analysis of significantly changed genes in E18.5 KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig3-figsupp3-data1-v2.xlsx
Figure 3—figure supplement 4
Expression of a collection of non-photoreceptor specific genes is largely unchanged in NMNAT1 knockout retinas.
Relative expression of indicated bipolar cell (A,D), amacrine and horizontal cell (B,E) and ganglion cell (C,F) genes in E18.5 (A–C) or P4 (D–F) knockout retinas as assessed by RNA-sequencing. Data are represented as mean ± SD. n = 3 biological replicates for (A–C), n = 5 biological replicates for (D–F).
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Figure 3—figure supplement 4—source data 1
Numerical source data for expression (relative to WT) of retinal cell-type-specific genes in E18.5 and P4 KO retinas from RNA-seq dataset.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig3-figsupp4-data1-v2.xlsx
Figure 3—figure supplement 5
NMNAT1-loss during retinal development affects formation of the outer plexiform layer.
(A) Representative hematoxylin and eosin (H&E)-stained retinal cross-sections from P4 knockout (B, B’) and control (A, A’) mice. Relative expression of indicated synaptic genes in P4 (C) and E18.5 (D) knockout retinas as assessed by RNA-sequencing are also shown. (E–H) Representative retinal sections from knockout (-/-) and floxed littermate control (+/+) mice at the indicated ages labelled with antibodies against synaptophysin (SYPH). Data are represented as mean ± SD. n = 5 biological replicates for (C), n = 3 biological replicates for (D,E–H). Corresponding zoom panels are indicated with dotted rectangles. Scale bars, 30 μm.
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Figure 3—figure supplement 5—source data 1
Numerical source data for expression (relative to WT) of several synapse-specific genes in E18.5 and P4 KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig3-figsupp5-data1-v2.xlsx
Figure 4 with 2 supplements
NMNAT1 loss causes activation of multiple cell death pathways in the retina.
(A–D) Representative retinal sections from knockout (-/-) and floxed littermate control (+/+) mice at the indicated ages labelled with an antibody against active Caspase-3 (AC3). Corresponding zoom panels are indicated with dotted rectangles. Arrows denote pyknotic, AC3-negative nuclei. (E) Quantification of pyknotic nuclei in sections from knockout and control mice at the indicated ages, grouped by presence (AC3+) or absence (AC3-) of active Caspase-3 labeling. Relative expression of several apoptotic (F), pyroptotic (G), and necroptotic (H) genes in P4 knockout retinas as assessed by RNA-sequencing. (I) Relative expression of Sarm1 in P4 knockout and control retinas as assessed by RNA-sequencing. (J) Relative abundance of cyclic-ADP-ribose (cADPR) in P4 knockout and control retinas as measured by mass spectrometry (grey bars represent means). Data are represented as mean ± SD. Significance determined using unpaired t-tests for (E) and (J) or DESeq2 for (F–I) (see Materials and methods). n = 3 biological replicates per condition for (A–E), n = 5 biological replicates for (F–I), n = 6 biological replicates (one outlier removed) for (J). Scale bars, 30 μm.
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Figure 4—source data 1
Numerical source data for quantification of Caspase-3-positive (AC3+) and Caspase-3-negative (AC3-) pyknotic cells in WT and KO retinal sections.
Numerical source data for expression (relative to WT) of cell death-related genes in P4 KO retinas. Numerical source data for relative abundance of cyclic ADP-ribose (cADPR) in P4 WT and KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig4-data1-v2.xlsx
Figure 4—figure supplement 1
Deregulation of several cell death pathways in NMNAT1-null retinas preceding gross degeneration.
Relative expression of several apoptotic (A), pyroptotic (B), and necroptotic (C) genes in E18.5 knockout retinas as assessed by RNA-sequencing. (D) Relative expression of Sarm1 in E18.5 knockout and control retinas as assessed by RNA-sequencing. (E) relative abundance of cyclic-ADP-ribose (cADPR) in E18.5 knockout and control retinas as measured by mass spectrometry (grey bars represent means). (F) Western blot of retinal lysate from P4 knockout and control retinas stained with an antibody against gasdermin D (GSDMD) and tubulin loading control. Data are represented as mean ± SD. n = 5 biological replicates for (A–D), n = 3 biological replicates for (E). Significance determined using DeSeq2 (see Materials and methods) (A–D) or Student’s t-test (E).
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Figure 4—figure supplement 1—source data 1
Numerical source data for expression (relative to WT) of cell death-related genes in E18.5 KO retinas.
Numerical source data for relative abundance of cyclic ADP-ribose (cADPR) in E18.5 WT and KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig4-figsupp1-data1-v2.xlsx
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Figure 4—figure supplement 1—source data 2
Uncropped western blot of P4 KO and WT retinal lysate stained with anti-Gasdermin D (GSDMD) antibody.
Arrow denotes bands corresponding to GSDMD.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig4-figsupp1-data2-v2.zip
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Figure 4—figure supplement 1—source data 3
Uncropped western blot of P4 KO and WT retinal lysate stained with anti-beta tubulin antibody.
Arrow denotes bands corresponding to beta-tubulin.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig4-figsupp1-data3-v2.zip
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Figure 4—figure supplement 1—source data 4
Raw western blot scan of P4 WT and KO retinal lysate stained with anti-Gasdermin D (GSDMD) antibody (700 channel).
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig4-figsupp1-data4-v2.zip
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Figure 4—figure supplement 1—source data 5
Raw western blot scan of P4 WT and KO retinal lysate stained with anti-beta tubulin antibody (800 channel).
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig4-figsupp1-data5-v2.zip
Figure 4—figure supplement 2
Transcriptional survey of cell death in the NMNAT1-null retina.
Heatmaps depicting log-scaled expression (relative to average of control samples) of genes related to apoptosis (A,B), necroptosis (C), ferroptosis (D), and pyroptosis (E) in P4 knockout and control retina as assessed by RNA-sequencing. Asterisks depict statistical significance as determined using DESeq2 (see Materials and methods). n = 5 biological replicates.
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Figure 4—figure supplement 2—source data 1
Numerical source data for expression of cell death pathway genes in P4 WT and KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig4-figsupp2-data1-v2.xlsx
Figure 5 with 2 supplements
Loss of NMNAT1 impairs retinal central carbon metabolism.
(A) Schematic illustrating metabolomics experimental approach. (B) Results of metabolite set enrichment analysis (MSEA) on significantly changed metabolites in P4 knockout retinas. (C) Relative abundance of NAD+ pathway metabolites in P4 knockout retinas as assessed by mass spectrometry. (D) Schematic illustrating the major mammalian NAD+ synthesis pathways colored according to metabolite changes in (C); NMNAT1-catalyzed steps are indicated with asterisks. (E) Relative abundance of glycolysis, TCA cycle, and creatine metabolites in P4 knockout retinas as assessed by mass spectrometry. (F) Schematic depicting abbreviated glycolysis/TCA cycle pathway colored according to metabolite changes in (E); aldolase-catalyzed step is indicated by a single asterisk, while GAPDH-catalyzed step is marked with a double asterisk. (G) Heatmap of log-transformed relative expression of a set of glycolysis/TCA cycle genes in P4 knockout (-/-) and control retinas (+/+) as assessed by RNA-sequencing. (H) Relative abundance of pentose-phosphate pathway (PPP) metabolites in P4 knockout retinas as assessed by mass spectrometry. Data are represented as mean ± SD. n = 6 biological replicates for (B,C,E,H), n = 5 biological replicates for (G). Statistical significance in panels (C), (E), and (H) is denoted according to the boxed legend.
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Figure 5—source data 1
Metabolic pathway overrepresentation analysis on significantly changed metabolites in P4 KO retinas.
Numerical source data for abundance (relative to WT) of various metabolites in P4 KO retinas. Numerical source data for expression of glycolysis/TCA pathway enzymes in P4 WT and KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig5-data1-v2.xlsx
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Figure 5—source data 2
Numerical source data for E18.5 and P4 LC-MS/MS experiments; peak intensity tables for all detected metabolites in E18.5 and P4 KO and WT retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig5-data2-v2.xlsx
Figure 5—figure supplement 1
Additional metabolic changes in NMNAT1-null retinas.
(A) Relative abundance of indicated carnitine species in P4 knockout retinas as assessed by mass spectrometry. (B) Relative expression of Aldh8a1 and Ogdhl in E18.5 knockout retinas as measured by RNA-sequencing. Principal component analysis (PCA) of E18.5 (C) and P4 (D) metabolomics data reveals expected clustering by genotype. Data are represented as mean ± SD. n = 6 biological replicates for (A), n = 5 biological replicates for (B).
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Figure 5—figure supplement 1—source data 1
Numerical source data for abundance (relative to WT) of acylcarnitine species in P4 KO retinas.
Numerical source data for expression (relative to WT) of Aldh8a1 and Ogdhl in E18.5 KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig5-figsupp1-data1-v2.xlsx
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Figure 5—figure supplement 1—source data 2
Numerical source data for abundance (relative to WT) of nucleotide and amino acid metabolites in P4 KO retinas.
- https://cdn.elifesciences.org/articles/71185/elife-71185-fig5-figsupp1-data2-v2.xlsx
Figure 5—figure supplement 2
NMNAT1-associated changes in retinal nucleotide and amino acid metabolism.
(A) Relative abundance of indicated nucleotide metabolites in P4 knockout retinas as assessed by mass spectrometry. (B) Relative abundance of indicated amino acid metabolites in P4 knockout retinas as assessed by mass spectrometry. Data are represented as mean ± SD. n = 6 biological replicates.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Mus musculus) | Nmnat1 | MGI:1913704 | ||
| Genetic reagent (mouse, male and female) | Nmnat1fl/fl | Conforti et al., 2011 | Nmnat1 gene exons 1 and 2 floxed | |
| Genetic reagent (mouse, male and female) | Six3-Cre | Christiansen et al., 2011 | Expresses Cre in developing eye bud (~E12.5) | |
| Cell line (Homo- sapiens) | HEK293T | ATCC | Cat# CRL-3216 | |
| Transfected construct (M. musculus) | FLAG-NMNAT1 | Genscript | Cat# OMu17664D | FLAG-tagged mouse NMNAT1 construct |
| Transfected construct (M. musculus) | FLAG-NMNAT2 | Genscript | Cat# OMu16562D | FLAG-tagged mouse NMNAT2 construct |
| Antibody | Anti-NMNAT1 (rabbit polyclonal) | this paper | WB (1:1000)See Materials and methods | |
| Antibody | Anti-beta tubulin (mouse monoclonal) | Sigma | Cat# T5201,RRID: AB_609915 | WB (1:3000) |
| Antibody | Anti-FLAG (mouse monoclonal) | Sigma | Cat# F1804,RRID: AB_262044 | WB (1:2000) |
| Antibody | Anti-BRN3A (mouse monoclonal) | Santa Cruz | Cat#: sc-8429,RRID: AB_626765 | IF (1:150) |
| Antibody | Anti-calretinin (rabbit polyclonal) | Millipore Sigma | Cat# AB5054RRID: AB_2068506 (discontinued) | IF (1:500) |
| Antibody | Anti-CHX10 (mouse monoclonal) | Santa Cruz | Cat# sc-365519,RRID: AB_10842442 | IF (1:50) |
| Antibody | Anti-recoverin (rabbit polyclonal) | Sigma | Cat# AB5585, RRID: AB_2253622 | IF (1:500) |
| Antibody | Anti-rhodopsin (4D2) (mouse monoclonal) | R.Molday, Univ. British Columbia | IF (1:500) | |
| Antibody | Anti-synaptophysin (rabbit monoclonal) | Thermo Fisher | Cat# MA5-14532, RRID: AB_10983675 | IF (1:1000) |
| Antibody | Anti-active caspase 3 (rabbit polyclonal) | R&D Systems | Cat# AF835, RRID: AB_2243952 | IF (1:500) |
| Antibody | Anti-OTX2 (rabbit polyclonal) | ProteinTech | Cat# 13497–1-AP, RRID: AB_2157176 | IF (1:500) |
| Antibody | Anti-phospho histone H3 (rabbit polyclonal) | Cell Signaling Technologies | Cat# 9701, RRID: AB_331535 | IF (1:1000) |
| Antibody | Anti-phosducin (sheep polyclonal) | Sokolov et al., 2004 | IF (1:1000) | |
| Antibody | Anti-M opsin (rabbit polyclonal) | Millipore Sigma | Cat# AB5045, RRID: AB_177456 | IF (1:1000) |
| Antibody | Anti-calbindin (mouse monoclonal) | Swant | Cat# 300, RRID: AB_10000347 | IF (1:500) |
| Antibody | Anti-rabbit Alexa Fluor- 568 (goat polyclonal) | Invitrogen | Cat# A-11011, RRID: AB_143157 | IF (1:1000) |
| Antibody | Anti-mouse Alexa Fluor- 488 (goat polyclonal) | Invitrogen | Cat# A-11001, RRID: AB_2534069 | IF (1:1000) |
| Antibody | Anti-rabbit Alexa Fluor 680 (goat polyclonal) | Invitrogen | Cat# A-21076, RRID: AB_2535736 | WB (1:50,000) |
| Antibody | Anti-mouse DyLight 800 | Invitrogen | Cat# SA5-10176, RRID: AB_2556756 | WB (1:50,000) |
| Antibody | Fluorescein Avidin D | Vector Laboratories | Cat# A-2001–5 | IF (1:500)For visualizing biotinylated PNA stain (below) |
| Other | Biotinylated peanut agglutinin (PNA) | Vector Laboratories | Cat# B-1075–5 | IF (1:500) |
| Other | 4′,6-diamidino-2-phenylindole (DAPI) | Thermo Fisher | Cat# D1306, RRID: AB_2629482 | IF (1:2000) |
| Software, algorithm | Fiji | Schindelin et al., 2012 | RRID:SCR_002285 | |
| Software, algorithm | FastQC | RRID:SCR_014583 | https://www.bioinformatics.babraham.ac.uk/projects/fastqc/ | |
| Software, algorithm | Bbduk | RRID:SCR_016969 | http://sourceforge.net/projects/bbmap/ | |
| Software, algorithm | HISAT2 2.1.0 | Kim et al., 2015 | RRID:SCR_015530 | |
| Software, algorithm | StringTie 1.3.6 | Pertea et al., 2015; Pertea et al., 2016 | RRID:SCR_016323 | |
| Software, algorithm | DeSeq2 R Package | Love et al., 2014 | RRID:SCR_015687 |
Additional files
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Supplementary file 1
Primer sequences for genotyping and RT-qPCR experiments and Mass spectrometry standards and parameters.
- https://cdn.elifesciences.org/articles/71185/elife-71185-supp1-v2.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/71185/elife-71185-transrepform1-v2.pdf
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Nuclear NAD+-biosynthetic enzyme NMNAT1 facilitates development and early survival of retinal neurons
eLife 10:e71185.
https://doi.org/10.7554/eLife.71185
