Journal: Cell Stem Cell

Article Title: Chromatin and Single-Cell RNA-Seq Profiling Reveal Dynamic Signaling and Metabolic Transitions during Human Spermatogonial Stem Cell Development

doi: 10.1016/j.stem.2017.09.003

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Article Snippet: Rabbit polyclonal anti-SPOCD1; dilution: 1:300 , Atlas Antibodies , cat# HPA031715, RRID: AB_2674008.

Techniques: Immunofluorescence, Histopathology, Methylation Sequencing, Software

Journal: Cell Death Discovery

Article Title: IFNγ regulates ferroptosis in KFs by inhibiting the expression of SPOCD1 through DNMT3A

doi: 10.1038/s41420-024-02257-z

Figure Lengend Snippet: A , B The volcano and heatmap plot illustrate the differentially expressed genes related to fibroblasts’ activities of the keloid group compared to the control group. C The GO function enrichment analysis of these differentially expressed genes. D , E Immunofluorescence staining and quantification of SPOCD1 in KD and NS (control group). F Western blotting assay of SPOCD1 in KFs and NFs. Bar=100 μm. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Si-RNA containing SPOCD1, IFNγ, DNMT1, DNMT3A and DNMT3B (GenePharma, Shanghai, China) were diluted into 200 pmol using 250 mL serum-free Opti-MEM.

Techniques: Control, Immunofluorescence, Staining, Western Blot

Journal: Cell Death Discovery

Article Title: IFNγ regulates ferroptosis in KFs by inhibiting the expression of SPOCD1 through DNMT3A

doi: 10.1038/s41420-024-02257-z

Figure Lengend Snippet: A the cck-8 assay of KFs after treated with si-SPOCD1 and erastin(5 μM) for 72 h. B – D the gene expression of TGF-β1, col1 and col3 of KFs after treated with si-SPOCD1 and erastin (5 μM) for 24 h. E Fluorescence intensity detection of ROS of KFs after treated with si-SPOCD1 and erastin(5 μM) for 24 h. F Flow analysis of ROS positive cells of KFs after treated with si-SPOCD1 and erastin (5 μM) for 24 h. G Quantitative statistics of ROS positive cells. H Quantitative statistics of MDA. Bar = 100 μm.* p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Si-RNA containing SPOCD1, IFNγ, DNMT1, DNMT3A and DNMT3B (GenePharma, Shanghai, China) were diluted into 200 pmol using 250 mL serum-free Opti-MEM.

Techniques: CCK-8 Assay, Gene Expression, Fluorescence

Journal: Cell Death Discovery

Article Title: IFNγ regulates ferroptosis in KFs by inhibiting the expression of SPOCD1 through DNMT3A

doi: 10.1038/s41420-024-02257-z

Figure Lengend Snippet: A Heatmap of hierarchical clustering analysis for samples according to DMPs. B The distribution of keloid’s DNA methylation sites. C The GO function enrichment analysis of these differentially expressed genes regulated by DNA methylation, showing the top 16 enrichment results. D KEGG enrichment results for significantly expressed genes with altered DNA methylation in keloid, showing the top 15 enrichment results. E The CCK-8 assay of KFs after treated with SGI-1027(5 μM) for 5 h. F The expression of protein SPOCD1 and DNMT3A after treated with si-DNMT3A or oe-DNMT3A for 72 h. G Western blotting results of DNMT3A in KFs. H Ion strength of protein mass spectrometry analysis. I The ion strength of SPOCD1 as determined by mass spectrometry analysis. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Si-RNA containing SPOCD1, IFNγ, DNMT1, DNMT3A and DNMT3B (GenePharma, Shanghai, China) were diluted into 200 pmol using 250 mL serum-free Opti-MEM.

Techniques: DNA Methylation Assay, CCK-8 Assay, Expressing, Western Blot, Mass Spectrometry

Journal: Cell Death Discovery

Article Title: IFNγ regulates ferroptosis in KFs by inhibiting the expression of SPOCD1 through DNMT3A

doi: 10.1038/s41420-024-02257-z

Figure Lengend Snippet: A The results of wound healing. B Quantitative analysis of wound healing model. C – J Gene expression of SPOCD1, IFNγ, col1, col3, GPX4, GSH, SLC7A11 and SLC3A2 in KFs treated with si-IFNγ, si-SPOCD1 or si-IFNγ + SPOCD1 for 48 h. Bar=100 μm. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Si-RNA containing SPOCD1, IFNγ, DNMT1, DNMT3A and DNMT3B (GenePharma, Shanghai, China) were diluted into 200 pmol using 250 mL serum-free Opti-MEM.

Techniques: Gene Expression

Journal: Cell Death Discovery

Article Title: IFNγ regulates ferroptosis in KFs by inhibiting the expression of SPOCD1 through DNMT3A

doi: 10.1038/s41420-024-02257-z

Figure Lengend Snippet: A Fluorescence intensity detection of ROS of KFs after treated with si-IFNγ, si-SPOCD1 or si-IFNγ + SPOCD1 for 48 h. B Flow analysis of ROS of KFs after treated with si-IFNγ, si-SPOCD1 or si-IFNγ + SPOCD1 for 48 h. C Quantitative statistics of ROS positive rate. D ) Electron microscopy detection results of KFs treated with si-IFNγ, si-SPOCD1 or si-IFNγ + SPOCD1 for 72 h. E The western blotting results of KFs after treated with si-IFNγ, si-SPOCD1 or si-IFNγ + SPOCD1 for 72 h. Blue bar=2 μm, Green bar=500 nm, Orange bar=50 μm.* p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Si-RNA containing SPOCD1, IFNγ, DNMT1, DNMT3A and DNMT3B (GenePharma, Shanghai, China) were diluted into 200 pmol using 250 mL serum-free Opti-MEM.

Techniques: Fluorescence, Electron Microscopy, Western Blot

Journal: Cell Death Discovery

Article Title: IFNγ regulates ferroptosis in KFs by inhibiting the expression of SPOCD1 through DNMT3A

doi: 10.1038/s41420-024-02257-z

Figure Lengend Snippet: Knockdown IFNγ induces a decrease in DNMT3A, which in turn increase SPOCD1 expression in KFs, then leads to col1, col3 excess and ROS reduction, and ultimately decreases KFs ferroptosis and ECM deposition. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Si-RNA containing SPOCD1, IFNγ, DNMT1, DNMT3A and DNMT3B (GenePharma, Shanghai, China) were diluted into 200 pmol using 250 mL serum-free Opti-MEM.

Techniques: Knockdown, Expressing