Western Blot:
Membrane:Article Title: Short-term methionine starvation reshapes diurnal hepatic m6A RNA methylation.
Article Snippet: .. Proteins were separated on 383 4-12% Bis-Tris Nupage gels (Invitrogen, NP0336BOX) and then transferred onto PVDF 384 membrane (Bio-Rad, 162-0218) followed by incubation with primary antibodies against MAT2A 385 (Abcam, ab186129), GAPDH (Santa Cruz, sc-32233) and CBS (Santa Cruz, sc-133154). .. Blots 386 were visualized using the ChemiDoc XRS system (Bio-Rad).
Incubation:Article Title: Short-term methionine starvation reshapes diurnal hepatic m6A RNA methylation.
Article Snippet: .. Proteins were separated on 383 4-12% Bis-Tris Nupage gels (Invitrogen, NP0336BOX) and then transferred onto PVDF 384 membrane (Bio-Rad, 162-0218) followed by incubation with primary antibodies against MAT2A 385 (Abcam, ab186129), GAPDH (Santa Cruz, sc-32233) and CBS (Santa Cruz, sc-133154). .. Blots 386 were visualized using the ChemiDoc XRS system (Bio-Rad).
Article Title: Deletion of α7 Nicotinic Acetylcholine Receptor in POMC Neurons Impairs Energy Balance and Leptin-Induced Anorexigenic Response in Male Mice.
Article Snippet: .. Nitrocellulose membranes were incubated overnight at 4 ◦C with specific antibodies such as pIRSSer307 (#2381; 1:1000), pSTAT3Tyr705 (#9145S; 1:1000), UCP1 (#14670; 1:1000), pAMPKThr172 (#2535S; 1:1000), pFOXO1Thr24/3Thr32 (#9464; 1:1000), pHSLSer565 (#4137; 1:1000), pAKTSer473 (#4060; 1:2000), pCREBSer133 (#9198; 1:1000), PI3K (#4292; 1:1000), PPAR- γ (#2443; 1:1000), Sirtuin 1 (#2310S; 1:1000), and vinculin (#139015; 1:2000) (Cell Signaling, Danvers, MA, USA), CPT2 (sc-377294; 1:1000), G6Pase (sc-25840; 1:1000), PEPCK (sc166778; 1:1000), GAPDH (sc-47724; 1:1000) (Santa Cruz Technology, Dallas, TX, USA), α-Tubulin (T5168; 1:5000) (Sigma-Aldrich, MI, USA), and Acetyl-STAT3Lys685 (PA5-17429; 1:1000) (Thermo Fisher, MA, USA). ..
Control:Article Title: Disease- and Region-Specific Molecular Patterns Involving miRNAs and Associated Signalling Pathways and Epithelial Cell-Fate Responses in PSC-Associated Ulcerative Colitis.
Article Snippet: Membranes were subsequently incubated with the appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies: anti-mouse IgG (1:5000; 115-035-146, Jackson ImmunoResearch Laboratories, West Grove, PA, USA) or anti-rabbit IgG (1:5000; BA1054, Boster Biological Technology, Pleasanton, CA, USA). .. GAPDH (1:5000; sc-47724 HRP, Santa Cruz Biotechnology) was utilised as a loading control. .. Immunoreactive bands were then subjected to visualisation using a chemiluminescent HRP substrate (Millipore, Burlington, MA, USA).
other:Article Title: EGFR activity governs stage-dependent fibrotic responses in recessive dystrophic epidermolysis bullosa.
Article Snippet: Primary antibody Clone/Code Company Dilution COL7A1 NC2-10 In house (Bornert et al, 2016) WB 1:1,000 β-tubulin 2-28-33/T-5239 Sigma-Aldrich WB 1:1,000 αSMA 1A4/A5228 Sigma-Aldrich WB 1:1,000; IF 1:500 TNC 578/MAB2138 R&D Systems WB 1:500 TSP1 A6.1/sc-59887 Santa Cruz WB 1:1,000 COL12A1 ab121304 Abcam WB 1:1,000 pSMAD2 138D4/3108S Cell Signaling WB 1:1,000 pSMAD3 EP823Y/ab52903 Abcam WB 1:1,000 pSMAD2/3 Ser 423,425/sc- 11769 Santa Cruz WB 1:1,000 EGFR pAb/NB100-596 Novus WB 1:1,000; IF 1:200 EGFR DAK-H1- WT/M7298 DAKO IF 1:50 pEGFR EP774Y/ab40815 Abcam WB 1:1,000; IF 1:500 pAKT D9E/4060S Cell Signaling WB 1:1,000 AKT 9272 Cell Signaling WB 1:1,000 pSTAT3 D3A7/9145S Cell Signaling WB 1:1,000 ADAM17 JM10-35/MA5- 32572 Thermo Fisher WB 1:1,000; IF 1:100 Jo urn al Pr e-p roo f 15 CAV1 pAb/ab2910 Abcam WB 1:1,000; IF 1:500 GAPDH 0411/sc-47724 Santa Cruz WB 1:1,000 FN pAb/ab2413 Abcam WB 1:1,000; IF 1:500 ERK pAb/M5670 Sigma-Aldrich WB 1:2,000 pERK MAPK- YT/M8159 Sigma-Aldrich WB 1:1,000 Abbreviations: pAb, polyclonalantibody Secondary antibody Product code Company Dilution HRP- conjugated anti-rabbit 5450-0010 SeraCare 1:10,000 HRP- conjugated anti-mouse 401253 Sigma-Aldrich 1:5,000 HRP- conjugated anti-rat 112-035-167 Jackson Immuno 1:10,000 Goat anti-rabbit conjugated to Alexa Fluor 488 A11070 Thermo Fisher 1:1,000 Goat anti-mouse conjugated to Alexa Fluor 488 A11029 Thermo Fisher 1:1,000 Goat anti-rabbit conjugated to Alexa Fluor 594 A11012 Thermo Fisher 1:1,000 Goat anti-mouse conjugated to Alexa Fluor 594 A11020 Thermo Fisher 1:1,000 Jo urn al Pr e-p roo f 16
Cell-Signaling:Article Title: Deletion of α7 Nicotinic Acetylcholine Receptor in POMC Neurons Impairs Energy Balance and Leptin-Induced Anorexigenic Response in Male Mice.
Article Snippet: .. Nitrocellulose membranes were incubated overnight at 4 ◦C with specific antibodies such as pIRSSer307 (#2381; 1:1000), pSTAT3Tyr705 (#9145S; 1:1000), UCP1 (#14670; 1:1000), pAMPKThr172 (#2535S; 1:1000), pFOXO1Thr24/3Thr32 (#9464; 1:1000), pHSLSer565 (#4137; 1:1000), pAKTSer473 (#4060; 1:2000), pCREBSer133 (#9198; 1:1000), PI3K (#4292; 1:1000), PPAR- γ (#2443; 1:1000), Sirtuin 1 (#2310S; 1:1000), and vinculin (#139015; 1:2000) (Cell Signaling, Danvers, MA, USA), CPT2 (sc-377294; 1:1000), G6Pase (sc-25840; 1:1000), PEPCK (sc166778; 1:1000), GAPDH (sc-47724; 1:1000) (Santa Cruz Technology, Dallas, TX, USA), α-Tubulin (T5168; 1:5000) (Sigma-Aldrich, MI, USA), and Acetyl-STAT3Lys685 (PA5-17429; 1:1000) (Thermo Fisher, MA, USA). ..
Article Title: Nervonic Acid Promotes Skin Regeneration by Enhancing Collagen Synthesis and Suppressing Inflammation
Article Snippet: .. The following primary antibodies were used: phospho‐p38, p38, phospho‐SMAD2/3, SMAD2/3, AKT, phospho‐mTOR, and mTOR (Cell Signaling Technology, Danvers, MA, USA); phospho‐AKT, phospho‐ERK, and ERK (R&D Systems, Minneapolis, MN, USA); COL1A1 (Novus Biologicals LLC, Centennial, CO, USA); α‐tubulin (Sigma‐Aldrich); and GAPDH (Santa Cruz Biotechnology, Dallas, TX, USA). .. Horseradish peroxidase‐conjugated anti‐rabbit and anti‐mouse IgG (AbClon, Seoul, Korea) were used as secondary antibodies.
Article Title: LncRNA Arsa-DT is required for spermatogenesis and male fertility by regulating G2/M progression.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Cyclin B1 Monoclonal Antibody Thermo Fisher Scientific Cat# MA1-155; RRID: AB_2536863 Anti-Cyclin D1 Monoclonal Antibody Abcam Cat# ab16663; RRID: AB_443423 Anti-Phospho-cdc2 (Tyr15) Antibody Cell Signaling Technology Cat# 9111; RRID: AB_331460 Anti-Phospho-Histone H3 (Ser10) Antibody Cell Signaling Technology Cat# 9701; RRID: AB_331535 Anti-Caspase-3 Antibody Cell Signaling Technology Cat# 9665; RRID: AB_2069872 Anti-PARP Antibody Cell Signaling Technology Cat# 9532; RRID: AB_659884 Anti-GAPDH Antibody Santa Cruz Biotechnology Cat# sc-32233; RRID: AB_627679 Rabbit anti-Goat IgG (H + L) Cross-Adsorbed Secondary Antibody, HRP Thermo Fisher Scientific Cat# R-21459; RRID: AB_2556529 Goat anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, HRP Thermo Fisher Scientific Cat# G-21040; RRID: AB_2536527 Anti-Ki67 Antibody Abcam Cat# ab16667; RRID: AB_302459 Anti-Histone H2A.X (phospho S139) antibody Abcam Cat# ab81299; RRID: AB_1640564 Anti-alpha-Tubulin Antibody Millipore Cat# 05-829; RRID: AB_310035 Anti-CD117/c-kit Antibody R&D systems Cat# AF1356; RRID: AB_354750 Anti-PLZF Antibody R&D systems Cat# AF2944; RRID: AB_2218943 Anti-STAR Antibody ProteinTech Cat# 80751-1; RRID: AB_2918910 Anti-SOX9 Antibody Thermo Fisher Scientific Cat# 14-9765-82; RRID: AB_2573006 Chicken Anti-Rabbit IgG (H + L) Antibody, Alexa FluorTM 488 Conjugated Thermo Fisher Scientific Cat# A-21441; RRID: AB_2535859 Chicken Anti-Mouse IgG (H + L) Antibody, Alexa FluorTM 488 Conjugated Thermo Fisher Scientific Cat# A-21200; RRID: AB_2535786 Donkey Anti-Goat IgG (H + L) Antibody, Alexa FluorTM 488 Conjugated Thermo Fisher Scientific Cat# A-11055; RRID: AB_2534102 Donkey Anti-Rabbit IgG (H + L) Polyclonal Antibody, Alexa FluorTM 555 Conjugated Thermo Fisher Scientific Cat# A-31572; RRID: AB_162543 Goat Anti-Mouse IgG (H + L) Highly Cross-adsorbed Antibody, Alexa FluorTM 633 Conjugated Thermo Fisher Scientific Cat# A-21052; RRID: AB_2535719 Goat Anti-Rabbit IgG (H + L) Highly Cross-adsorbed Antibody, Alexa FluorTM 647 Conjugated Thermo Fisher Scientific Cat# A-21245; RRID: AB_2535813 Anti-CD45 Antibody, anti-mouse, Biotin Miltenyi Biotec Cat# 130-124-209; RRID: AB_2819580 Anti-CD16/CD32 Antibody, anti-mouse, Biotin Miltenyi Biotec Cat# 130-101-895; RRID: AB_2660226 Anti-CD326 (EpCAM) Monoclonal Antibody (G8.8), PE Thermo Fisher Scientific Cat# 12-5791-81; RRID: AB_953619 Anti-phospho-Ser/Thr-Pro, MPM-2 Antibody Millipore Cat# 05-368; RRID: AB_309698 Chemicals, peptides, and recombinant proteins Dulbecco’s modified Eagle’s medium Cytiva SH30243.01 Fetal bovine serum Cytiva SH31194.03 Penicillin-Streptomycin solution Cytiva SV30010 Lipofectamine 3000 Thermo Fisher Scientific L3000001 Nocodazole Sigma Aldrich SML1665 i-StarTaqTM DNA polymerase iNtRON Biotechnology 25161 TRIzolTM Reagent Thermo Fisher Scientific 15596026 Collagenase type IV Sigma Aldrich C5138 DNase I Sigma Aldrich DN25 IGEPAL CA-630 (NP-40) Sigma Aldrich I3021 (Continued on next page) iScience 29, 117813, November 20, 2026 13 .. REAGENT or RESOURCE SOURCE IDENTIFIER Easy-Blue phenol buffer iNtRON Biotechnology 17061 FITC-conjugated peanut agglutinin (PNA) Sigma Aldrich L7381 M-MLV Reverse Transcriptase Thermo Fisher Scientific 28025013 Donkey serum Sigma Aldrich D9663 Bovine serum albumin MP Biomedicals MP 0216006980 TritonTM X-100 Sigma Aldrich T9284 Dihydroethidium Sigma Aldrich 37291 Anti-PE MicroBeads Miltenyi Biotec 130-048-801 Streptavidin MicroBeads Miltenyi Biotec 130-048-102 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Sigma Aldrich 475989 Propidium iodide Sigma Aldrich P4170 cOmpleteTM, Mini Protease Inhibitor Cocktail Roche 11836153001 PhosSTOPTM Roche 4906845001 (PHOSS-RO) AmershamTM ECL Western Blotting Detection Reagent Cytiva RPN2105 AmershamTM ECL Prime Western Blotting Detection Reagent Cytiva RPN2232 NICSROWESTTM ECL PICO Bionics BNROW-0007 NICSROWESTTM ECL ADVANCE Bionics BNROP-0009 Critical commercial assays Testosterone ELISA Kit Cayman Chemical 582701 Periodic Acid Schiff (PAS) Stain Kit ScyTek Laboratories PAS-1 GeneRacerTM Kit with SuperScriptTM III RT and TOPO TA CloningTM Kit for Sequencing Thermo Fisher Scientific L150201 in situ Apoptosis Detection Kit Takara MK500 Rneasy Mini kit Qiagen 74104 RNAscopeTM Multiplex Fluorescent Reagent Kit v2 Advanced Cell Diagnostics 323100 Pierce BCA protein assay kit Thermo Fisher Scientific 23225 Deposited data Mouse testes single-cell RNA-seq data Yuan et al.19 GEO: GSE254812 Human testes RNA-seq data Balagannavar et al.29 GEO: GSE216907 Experimental models: Cell lines GC-1 spg ATCC CRL-2053 Experimental models: Organisms/strains C57BL/6 mice Orient Bio C57BL/6NCrlOri C57BL/6 Arsa-DT− /− mice In house N/A Oligonucleotides Arsa-DT (C230037L18Rik) primer In house Forward 5′- CCAGGACCTGCGCATTATT -3′; Reverse 5′- CCTCTTCGCTTGGTGTCTTT -3′ Stra8 primer In house Forward 5′- CTGTGGCAGGTACTGAATAGG -3′; Reverse 5′- GCAGGTTGAAGGATGCTTTG -3′ Arsa primer In house Forward 5′- CCTTGTTCCCATCCCACTAC -3′; Reverse 5′- GTCTCGGGAGAAAGACACATAC -3′ Shank3 primer In house Forward 5′- CCGATACAAGCGGAGAGTTTAT -3′; Reverse 5′- CCACCTTATCTGTGCTGTGTAG -3′ Gapdh primer In house Forward 5′- GTGGCAAAGTGGAGATTGTTG -3′; Reverse 5′- CGTTGAATTTGCCGTGAGTG -3′ Arsa-DT Transcript 1 primer In house Forward 5′- ACATTCCAGCACGGTTTCTG -3′; Reverse 5′- GTGTGGGTGTTTTACCTGCATTA -3′ (Continued on next page) 14 iScience 29, 117813, November 20, 2026
Recombinant:Article Title: LncRNA Arsa-DT is required for spermatogenesis and male fertility by regulating G2/M progression.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Cyclin B1 Monoclonal Antibody Thermo Fisher Scientific Cat# MA1-155; RRID: AB_2536863 Anti-Cyclin D1 Monoclonal Antibody Abcam Cat# ab16663; RRID: AB_443423 Anti-Phospho-cdc2 (Tyr15) Antibody Cell Signaling Technology Cat# 9111; RRID: AB_331460 Anti-Phospho-Histone H3 (Ser10) Antibody Cell Signaling Technology Cat# 9701; RRID: AB_331535 Anti-Caspase-3 Antibody Cell Signaling Technology Cat# 9665; RRID: AB_2069872 Anti-PARP Antibody Cell Signaling Technology Cat# 9532; RRID: AB_659884 Anti-GAPDH Antibody Santa Cruz Biotechnology Cat# sc-32233; RRID: AB_627679 Rabbit anti-Goat IgG (H + L) Cross-Adsorbed Secondary Antibody, HRP Thermo Fisher Scientific Cat# R-21459; RRID: AB_2556529 Goat anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, HRP Thermo Fisher Scientific Cat# G-21040; RRID: AB_2536527 Anti-Ki67 Antibody Abcam Cat# ab16667; RRID: AB_302459 Anti-Histone H2A.X (phospho S139) antibody Abcam Cat# ab81299; RRID: AB_1640564 Anti-alpha-Tubulin Antibody Millipore Cat# 05-829; RRID: AB_310035 Anti-CD117/c-kit Antibody R&D systems Cat# AF1356; RRID: AB_354750 Anti-PLZF Antibody R&D systems Cat# AF2944; RRID: AB_2218943 Anti-STAR Antibody ProteinTech Cat# 80751-1; RRID: AB_2918910 Anti-SOX9 Antibody Thermo Fisher Scientific Cat# 14-9765-82; RRID: AB_2573006 Chicken Anti-Rabbit IgG (H + L) Antibody, Alexa FluorTM 488 Conjugated Thermo Fisher Scientific Cat# A-21441; RRID: AB_2535859 Chicken Anti-Mouse IgG (H + L) Antibody, Alexa FluorTM 488 Conjugated Thermo Fisher Scientific Cat# A-21200; RRID: AB_2535786 Donkey Anti-Goat IgG (H + L) Antibody, Alexa FluorTM 488 Conjugated Thermo Fisher Scientific Cat# A-11055; RRID: AB_2534102 Donkey Anti-Rabbit IgG (H + L) Polyclonal Antibody, Alexa FluorTM 555 Conjugated Thermo Fisher Scientific Cat# A-31572; RRID: AB_162543 Goat Anti-Mouse IgG (H + L) Highly Cross-adsorbed Antibody, Alexa FluorTM 633 Conjugated Thermo Fisher Scientific Cat# A-21052; RRID: AB_2535719 Goat Anti-Rabbit IgG (H + L) Highly Cross-adsorbed Antibody, Alexa FluorTM 647 Conjugated Thermo Fisher Scientific Cat# A-21245; RRID: AB_2535813 Anti-CD45 Antibody, anti-mouse, Biotin Miltenyi Biotec Cat# 130-124-209; RRID: AB_2819580 Anti-CD16/CD32 Antibody, anti-mouse, Biotin Miltenyi Biotec Cat# 130-101-895; RRID: AB_2660226 Anti-CD326 (EpCAM) Monoclonal Antibody (G8.8), PE Thermo Fisher Scientific Cat# 12-5791-81; RRID: AB_953619 Anti-phospho-Ser/Thr-Pro, MPM-2 Antibody Millipore Cat# 05-368; RRID: AB_309698 Chemicals, peptides, and recombinant proteins Dulbecco’s modified Eagle’s medium Cytiva SH30243.01 Fetal bovine serum Cytiva SH31194.03 Penicillin-Streptomycin solution Cytiva SV30010 Lipofectamine 3000 Thermo Fisher Scientific L3000001 Nocodazole Sigma Aldrich SML1665 i-StarTaqTM DNA polymerase iNtRON Biotechnology 25161 TRIzolTM Reagent Thermo Fisher Scientific 15596026 Collagenase type IV Sigma Aldrich C5138 DNase I Sigma Aldrich DN25 IGEPAL CA-630 (NP-40) Sigma Aldrich I3021 (Continued on next page) iScience 29, 117813, November 20, 2026 13 .. REAGENT or RESOURCE SOURCE IDENTIFIER Easy-Blue phenol buffer iNtRON Biotechnology 17061 FITC-conjugated peanut agglutinin (PNA) Sigma Aldrich L7381 M-MLV Reverse Transcriptase Thermo Fisher Scientific 28025013 Donkey serum Sigma Aldrich D9663 Bovine serum albumin MP Biomedicals MP 0216006980 TritonTM X-100 Sigma Aldrich T9284 Dihydroethidium Sigma Aldrich 37291 Anti-PE MicroBeads Miltenyi Biotec 130-048-801 Streptavidin MicroBeads Miltenyi Biotec 130-048-102 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Sigma Aldrich 475989 Propidium iodide Sigma Aldrich P4170 cOmpleteTM, Mini Protease Inhibitor Cocktail Roche 11836153001 PhosSTOPTM Roche 4906845001 (PHOSS-RO) AmershamTM ECL Western Blotting Detection Reagent Cytiva RPN2105 AmershamTM ECL Prime Western Blotting Detection Reagent Cytiva RPN2232 NICSROWESTTM ECL PICO Bionics BNROW-0007 NICSROWESTTM ECL ADVANCE Bionics BNROP-0009 Critical commercial assays Testosterone ELISA Kit Cayman Chemical 582701 Periodic Acid Schiff (PAS) Stain Kit ScyTek Laboratories PAS-1 GeneRacerTM Kit with SuperScriptTM III RT and TOPO TA CloningTM Kit for Sequencing Thermo Fisher Scientific L150201 in situ Apoptosis Detection Kit Takara MK500 Rneasy Mini kit Qiagen 74104 RNAscopeTM Multiplex Fluorescent Reagent Kit v2 Advanced Cell Diagnostics 323100 Pierce BCA protein assay kit Thermo Fisher Scientific 23225 Deposited data Mouse testes single-cell RNA-seq data Yuan et al.19 GEO: GSE254812 Human testes RNA-seq data Balagannavar et al.29 GEO: GSE216907 Experimental models: Cell lines GC-1 spg ATCC CRL-2053 Experimental models: Organisms/strains C57BL/6 mice Orient Bio C57BL/6NCrlOri C57BL/6 Arsa-DT− /− mice In house N/A Oligonucleotides Arsa-DT (C230037L18Rik) primer In house Forward 5′- CCAGGACCTGCGCATTATT -3′; Reverse 5′- CCTCTTCGCTTGGTGTCTTT -3′ Stra8 primer In house Forward 5′- CTGTGGCAGGTACTGAATAGG -3′; Reverse 5′- GCAGGTTGAAGGATGCTTTG -3′ Arsa primer In house Forward 5′- CCTTGTTCCCATCCCACTAC -3′; Reverse 5′- GTCTCGGGAGAAAGACACATAC -3′ Shank3 primer In house Forward 5′- CCGATACAAGCGGAGAGTTTAT -3′; Reverse 5′- CCACCTTATCTGTGCTGTGTAG -3′ Gapdh primer In house Forward 5′- GTGGCAAAGTGGAGATTGTTG -3′; Reverse 5′- CGTTGAATTTGCCGTGAGTG -3′ Arsa-DT Transcript 1 primer In house Forward 5′- ACATTCCAGCACGGTTTCTG -3′; Reverse 5′- GTGTGGGTGTTTTACCTGCATTA -3′ (Continued on next page) 14 iScience 29, 117813, November 20, 2026
Modification:Article Title: LncRNA Arsa-DT is required for spermatogenesis and male fertility by regulating G2/M progression.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Cyclin B1 Monoclonal Antibody Thermo Fisher Scientific Cat# MA1-155; RRID: AB_2536863 Anti-Cyclin D1 Monoclonal Antibody Abcam Cat# ab16663; RRID: AB_443423 Anti-Phospho-cdc2 (Tyr15) Antibody Cell Signaling Technology Cat# 9111; RRID: AB_331460 Anti-Phospho-Histone H3 (Ser10) Antibody Cell Signaling Technology Cat# 9701; RRID: AB_331535 Anti-Caspase-3 Antibody Cell Signaling Technology Cat# 9665; RRID: AB_2069872 Anti-PARP Antibody Cell Signaling Technology Cat# 9532; RRID: AB_659884 Anti-GAPDH Antibody Santa Cruz Biotechnology Cat# sc-32233; RRID: AB_627679 Rabbit anti-Goat IgG (H + L) Cross-Adsorbed Secondary Antibody, HRP Thermo Fisher Scientific Cat# R-21459; RRID: AB_2556529 Goat anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, HRP Thermo Fisher Scientific Cat# G-21040; RRID: AB_2536527 Anti-Ki67 Antibody Abcam Cat# ab16667; RRID: AB_302459 Anti-Histone H2A.X (phospho S139) antibody Abcam Cat# ab81299; RRID: AB_1640564 Anti-alpha-Tubulin Antibody Millipore Cat# 05-829; RRID: AB_310035 Anti-CD117/c-kit Antibody R&D systems Cat# AF1356; RRID: AB_354750 Anti-PLZF Antibody R&D systems Cat# AF2944; RRID: AB_2218943 Anti-STAR Antibody ProteinTech Cat# 80751-1; RRID: AB_2918910 Anti-SOX9 Antibody Thermo Fisher Scientific Cat# 14-9765-82; RRID: AB_2573006 Chicken Anti-Rabbit IgG (H + L) Antibody, Alexa FluorTM 488 Conjugated Thermo Fisher Scientific Cat# A-21441; RRID: AB_2535859 Chicken Anti-Mouse IgG (H + L) Antibody, Alexa FluorTM 488 Conjugated Thermo Fisher Scientific Cat# A-21200; RRID: AB_2535786 Donkey Anti-Goat IgG (H + L) Antibody, Alexa FluorTM 488 Conjugated Thermo Fisher Scientific Cat# A-11055; RRID: AB_2534102 Donkey Anti-Rabbit IgG (H + L) Polyclonal Antibody, Alexa FluorTM 555 Conjugated Thermo Fisher Scientific Cat# A-31572; RRID: AB_162543 Goat Anti-Mouse IgG (H + L) Highly Cross-adsorbed Antibody, Alexa FluorTM 633 Conjugated Thermo Fisher Scientific Cat# A-21052; RRID: AB_2535719 Goat Anti-Rabbit IgG (H + L) Highly Cross-adsorbed Antibody, Alexa FluorTM 647 Conjugated Thermo Fisher Scientific Cat# A-21245; RRID: AB_2535813 Anti-CD45 Antibody, anti-mouse, Biotin Miltenyi Biotec Cat# 130-124-209; RRID: AB_2819580 Anti-CD16/CD32 Antibody, anti-mouse, Biotin Miltenyi Biotec Cat# 130-101-895; RRID: AB_2660226 Anti-CD326 (EpCAM) Monoclonal Antibody (G8.8), PE Thermo Fisher Scientific Cat# 12-5791-81; RRID: AB_953619 Anti-phospho-Ser/Thr-Pro, MPM-2 Antibody Millipore Cat# 05-368; RRID: AB_309698 Chemicals, peptides, and recombinant proteins Dulbecco’s modified Eagle’s medium Cytiva SH30243.01 Fetal bovine serum Cytiva SH31194.03 Penicillin-Streptomycin solution Cytiva SV30010 Lipofectamine 3000 Thermo Fisher Scientific L3000001 Nocodazole Sigma Aldrich SML1665 i-StarTaqTM DNA polymerase iNtRON Biotechnology 25161 TRIzolTM Reagent Thermo Fisher Scientific 15596026 Collagenase type IV Sigma Aldrich C5138 DNase I Sigma Aldrich DN25 IGEPAL CA-630 (NP-40) Sigma Aldrich I3021 (Continued on next page) iScience 29, 117813, November 20, 2026 13 .. REAGENT or RESOURCE SOURCE IDENTIFIER Easy-Blue phenol buffer iNtRON Biotechnology 17061 FITC-conjugated peanut agglutinin (PNA) Sigma Aldrich L7381 M-MLV Reverse Transcriptase Thermo Fisher Scientific 28025013 Donkey serum Sigma Aldrich D9663 Bovine serum albumin MP Biomedicals MP 0216006980 TritonTM X-100 Sigma Aldrich T9284 Dihydroethidium Sigma Aldrich 37291 Anti-PE MicroBeads Miltenyi Biotec 130-048-801 Streptavidin MicroBeads Miltenyi Biotec 130-048-102 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Sigma Aldrich 475989 Propidium iodide Sigma Aldrich P4170 cOmpleteTM, Mini Protease Inhibitor Cocktail Roche 11836153001 PhosSTOPTM Roche 4906845001 (PHOSS-RO) AmershamTM ECL Western Blotting Detection Reagent Cytiva RPN2105 AmershamTM ECL Prime Western Blotting Detection Reagent Cytiva RPN2232 NICSROWESTTM ECL PICO Bionics BNROW-0007 NICSROWESTTM ECL ADVANCE Bionics BNROP-0009 Critical commercial assays Testosterone ELISA Kit Cayman Chemical 582701 Periodic Acid Schiff (PAS) Stain Kit ScyTek Laboratories PAS-1 GeneRacerTM Kit with SuperScriptTM III RT and TOPO TA CloningTM Kit for Sequencing Thermo Fisher Scientific L150201 in situ Apoptosis Detection Kit Takara MK500 Rneasy Mini kit Qiagen 74104 RNAscopeTM Multiplex Fluorescent Reagent Kit v2 Advanced Cell Diagnostics 323100 Pierce BCA protein assay kit Thermo Fisher Scientific 23225 Deposited data Mouse testes single-cell RNA-seq data Yuan et al.19 GEO: GSE254812 Human testes RNA-seq data Balagannavar et al.29 GEO: GSE216907 Experimental models: Cell lines GC-1 spg ATCC CRL-2053 Experimental models: Organisms/strains C57BL/6 mice Orient Bio C57BL/6NCrlOri C57BL/6 Arsa-DT− /− mice In house N/A Oligonucleotides Arsa-DT (C230037L18Rik) primer In house Forward 5′- CCAGGACCTGCGCATTATT -3′; Reverse 5′- CCTCTTCGCTTGGTGTCTTT -3′ Stra8 primer In house Forward 5′- CTGTGGCAGGTACTGAATAGG -3′; Reverse 5′- GCAGGTTGAAGGATGCTTTG -3′ Arsa primer In house Forward 5′- CCTTGTTCCCATCCCACTAC -3′; Reverse 5′- GTCTCGGGAGAAAGACACATAC -3′ Shank3 primer In house Forward 5′- CCGATACAAGCGGAGAGTTTAT -3′; Reverse 5′- CCACCTTATCTGTGCTGTGTAG -3′ Gapdh primer In house Forward 5′- GTGGCAAAGTGGAGATTGTTG -3′; Reverse 5′- CGTTGAATTTGCCGTGAGTG -3′ Arsa-DT Transcript 1 primer In house Forward 5′- ACATTCCAGCACGGTTTCTG -3′; Reverse 5′- GTGTGGGTGTTTTACCTGCATTA -3′ (Continued on next page) 14 iScience 29, 117813, November 20, 2026
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