Journal: Nature Communications

Article Title: Fibroblast-specific PRMT5 deficiency suppresses cardiac fibrosis and left ventricular dysfunction in male mice

doi: 10.1038/s41467-024-46711-z

Figure Lengend Snippet: a , b TGF-β induces WDR5/MLL1-mediated H3K4 trimethylation, which is inhibited by PRMT5. Primary cultured cardiac fibroblasts from neonatal rats were treated with or without TGF-β (10 ng/mL) in the presence or absence of MM102 a or EPZ015666 b for 6 h. Chromatin immunoprecipitation with anti-histone H3K4me3 antibody was performed, followed by quantitative PCR analysis of the Col1a1 and Acta2 promoter sites. IgG was used as a negative control. c – f Primary cultured cardiac fibroblasts were treated with MM102 at the indicated concentration and then treated with or without TGF-β (10 ng/mL). Gene expression levels of Col1a1 and Acta2 were quantified by qRT-PCR c . The protein expression level of α-SMA was determined by western blotting and quantified with ImageJ software d . Primary cultured cardiac fibroblasts were transfected with siRNA (siControl, siMLL1, or siWDR5) and then treated with or without TGF-β (10 ng/mL). The protein expression of α-SMA was quantified by western blotting e – f . Values are presented as mean ± SD. a , b , e , f n = 3 biologically independent samples; c n = 6 biologically independent samples; d n = 4 biologically independent samples. One-way ANOVA, followed by Tukey’s multiple comparison test (a, b, e, f) or Dunnett’s multiple comparison test versus TGF-β-treated group c , d . P values are indicated in each graph. Source data are provided as a Source Data file.

Article Snippet: The following primary antibodies were used: anti-PRMT5 rabbit monoclonal antibody (Cat#07-405, Merck, Tokyo, Japan) at 1:5000 dilution, anti-Smad3 rabbit monoclonal antibody (#9523, Cell Signaling Technology, Tokyo, Japan) at 1:2000 dilution, anti-WDR5 rabbit monoclonal antibody (#13105, Cell Signaling Technology) at 1:2000 dilution, anti-MLL1 rabbit monoclonal antibody (#14197, Cell Signaling Technology) at 1:2000 dilution, anti-α-SMA mouse monoclonal antibody (Cat#A5228, Sigma-Aldrich) at 1:5000 dilution, anti-HA-tag rabbit monoclonal antibody (Cat#M132-3, MBL Life Science) at 1:10000 dilution, anti-FLAG-tag rabbit monoclonal antibody (Cat#M185-3L, MBL Life Science) at 1:10000 dilution, anti-MGEA5 polyclonal antibody (Cat#14711-1-AP, Proteintech) at 1:5000 dilution, Anti-Histone H3 (trimethyl K4) rabbit monoclonal antibody (Cat#ab8580, Abcam) at 1:5000 dilution, and anti-β-actin mouse monoclonal clone AC-15 IgG (Cat#A1978, Sigma-Aldrich) at 1:10000 dilution.

Techniques: Cell Culture, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Negative Control, Concentration Assay, Gene Expression, Quantitative RT-PCR, Expressing, Western Blot, Software, Transfection, Comparison

Journal: Nature Communications

Article Title: Fibroblast-specific PRMT5 deficiency suppresses cardiac fibrosis and left ventricular dysfunction in male mice

doi: 10.1038/s41467-024-46711-z

Figure Lengend Snippet: In the promoter regions of fibrotic genes, PRMT5 interacts with Smad3 and symmetrically dimethylates histone H3R2, and WDR5/MLL1 subsequently induces H3K4 trimethylation. These histone methylations trigger fibrotic gene transcription in cardiac fibroblasts, contributing to the progression of heart failure.

Article Snippet: The following primary antibodies were used: anti-PRMT5 rabbit monoclonal antibody (Cat#07-405, Merck, Tokyo, Japan) at 1:5000 dilution, anti-Smad3 rabbit monoclonal antibody (#9523, Cell Signaling Technology, Tokyo, Japan) at 1:2000 dilution, anti-WDR5 rabbit monoclonal antibody (#13105, Cell Signaling Technology) at 1:2000 dilution, anti-MLL1 rabbit monoclonal antibody (#14197, Cell Signaling Technology) at 1:2000 dilution, anti-α-SMA mouse monoclonal antibody (Cat#A5228, Sigma-Aldrich) at 1:5000 dilution, anti-HA-tag rabbit monoclonal antibody (Cat#M132-3, MBL Life Science) at 1:10000 dilution, anti-FLAG-tag rabbit monoclonal antibody (Cat#M185-3L, MBL Life Science) at 1:10000 dilution, anti-MGEA5 polyclonal antibody (Cat#14711-1-AP, Proteintech) at 1:5000 dilution, Anti-Histone H3 (trimethyl K4) rabbit monoclonal antibody (Cat#ab8580, Abcam) at 1:5000 dilution, and anti-β-actin mouse monoclonal clone AC-15 IgG (Cat#A1978, Sigma-Aldrich) at 1:10000 dilution.

Techniques:

Journal: Cell Reports Medicine

Article Title: Targeting pancreatic cancer glutamine dependency confers vulnerability to GPX4-dependent ferroptosis

doi: 10.1016/j.xcrm.2025.101928

Figure Lengend Snippet:

Article Snippet: Rabbit anti-WDR5 , Proteintech , Cat# 15544-1-AP; RRID: AB_2257220.

Techniques: Virus, CRISPR, Recombinant, Protease Inhibitor, Lysis, Ligation, Reverse Transcription, SYBR Green Assay, Glo Assay, Sample Prep, DNA Extraction, Enzyme-linked Immunosorbent Assay, Multiple Displacement Amplification, Flow Cytometry, Software