Journal: Advanced Science

Article Title: LEDGF Binds H3R17me2a Promoting De Novo Nucleotide Biosynthesis in SETD2 Mutant Clear Cell Renal Cell Carcinoma

doi: 10.1002/advs.202416809

Figure Lengend Snippet: CARM1 is highly expressed in ccRCC and promotes proliferation. A,B) CARM1 is highly expressed in ccRCC patients and is correlated with poor prognosis on the UALCAN database. C) CARM1 and H3R17me2a are highly expressed in ccRCC cells such as 786‐O, A498, and Caki‐1. D,E) Immunofluorescence staining using the tissue microarray revealed that CARM1 is highly expressed in ccRCC tissue samples. Scale bar = 100 µm. (Tumor n = 79, Normal n = 79) F) CcRCC patients with advanced T stage have higher CARM1 expression, which may be associated with poor prognosis. (T1 n = 53, T2 n = 23, T3 n = 3) G,H) Immunofluorescence staining using the tissue microarray revealed that H3R17me2a level is higher in ccRCC tissue samples. Scale bar = 100 µm. (Tumor n = 79, Normal n = 79) I) CcRCC patients with advanced T stage have higher H3R17me2a level, which may be associated with poor prognosis. (T1 n = 53, T2 n = 23, T3 n = 3) J) The siRNA targeting CARM1 mRNA can effectively interfere with CARM1 and decreases H3R17me2a in ccRCC cells. K–N) Reduction of CARM1 and H3R17me2a significantly inhibited the proliferation of ccRCC cells, as demonstrated by CCK‐8 (K), colony formation (L) and EdU proliferation assays (M‐N). Scale bar = 100 µm. Data are shown as mean ± SD. ** p < 0.01, *** p < 0.001.

Article Snippet: The antibodies used in this study are as follows: LEDGF (Abcam, ab177159), CARM1 (CST, #3379), H3R17me2a (Acive motif, #39 710), PRMT6 (Abcam, ab271091), H3K36me3 (CST, #4909), PPAT (Proteintech, #15401‐1‐AP), PAICS (Proteintech, #12967‐1‐AP), GART (Proteintech, #13659‐1‐AP), ADSL (Proteintech, #15264‐1‐AP), ADSS2 (Proteintech, #16373‐1‐AP), Histone H3 (Proteintech, #17168‐1‐AP), Alpha Actin (Proteintech, #23660‐1‐AP), Flag (Abmart, #M20008), Goat Anti‐Rabbit IgG (H + L) (Proteintech,#SA00001‐2), and Goat Anti‐Mouse IgG (H + L) (Proteintech, #SA00001‐1).

Techniques: Immunofluorescence, Staining, Microarray, Expressing, CCK-8 Assay

Journal: Advanced Science

Article Title: LEDGF Binds H3R17me2a Promoting De Novo Nucleotide Biosynthesis in SETD2 Mutant Clear Cell Renal Cell Carcinoma

doi: 10.1002/advs.202416809

Figure Lengend Snippet: Construction of the H3R17me2a‐deficient cell lines. A,B) Validation of knockout efficiency of CARM1 and the reduction of H3R17me2a level in transduced pooled A498 cells (A) and 786‐O cells (B). C–E) Validation of knockout efficiency of CARM1 and the reduction of H3R17me2a level in monoclonal A498 cells (C) and 786‐O cells (D‐E). F,G) Knockdown of PRMT6 using siRNA has little effect on H3R17me2a level in A498 cells (F) and 786‐O cells (G). H) The combined knockdown of CARM1 and PRMT6 using siRNAs reduces H3R17me2a level in A498 cells; knockdown of PRMT6 in CARM1‐KO cells significantly decreased H3R17me2a in A498 cells. I) Knockdown of PRMT6 in CARM1‐KO cells significantly decreased H3R17me2a in A498 cells. J) The combination of CARM1 (MCE, EZM2302 and TP‐064) and PRMT6 (MCE, EPZ020411) inhibitors can significantly reduce H3R17me2a level in A498 cells.

Article Snippet: The antibodies used in this study are as follows: LEDGF (Abcam, ab177159), CARM1 (CST, #3379), H3R17me2a (Acive motif, #39 710), PRMT6 (Abcam, ab271091), H3K36me3 (CST, #4909), PPAT (Proteintech, #15401‐1‐AP), PAICS (Proteintech, #12967‐1‐AP), GART (Proteintech, #13659‐1‐AP), ADSL (Proteintech, #15264‐1‐AP), ADSS2 (Proteintech, #16373‐1‐AP), Histone H3 (Proteintech, #17168‐1‐AP), Alpha Actin (Proteintech, #23660‐1‐AP), Flag (Abmart, #M20008), Goat Anti‐Rabbit IgG (H + L) (Proteintech,#SA00001‐2), and Goat Anti‐Mouse IgG (H + L) (Proteintech, #SA00001‐1).

Techniques: Biomarker Discovery, Knock-Out, Knockdown

Journal: Advanced Science

Article Title: LEDGF Binds H3R17me2a Promoting De Novo Nucleotide Biosynthesis in SETD2 Mutant Clear Cell Renal Cell Carcinoma

doi: 10.1002/advs.202416809

Figure Lengend Snippet: Deficiency of LEDGF protects NKG mice against xenograft proliferation. A) Schematic diagram of subcutaneous tumor model in NKG mice in indicated treatment groups. All surviving mice were euthanized 8 weeks after tumor cell inoculation. B) Knock out of LEDGF effectively reduced the proliferation of xenografts in NKG mice. (n = 5) C) There was no significant difference in body weight between the two groups throughout the experiment. D–F) Elimination of LEDGF effectively reduced the volume (D‐E) and weight (F) of NKG mice xenografts. G) QRT‐PCR was used to demonstrate that decrease of LEDGF can reduce mRNA expression of PPAT, PAICS, GART, ADSL, and ADSS2 in xenograft tumors. H) The proliferation ability of xenografts in LEDGF‐KO group was significantly reduced. The expression levels of PPAT, PAICS, GART, and ADSL were significantly decreased, while ADSS2 expression was almost unchanged. Scale bar = 100 µm. I) A schematic model illustrating that LEDGF interacts with CARM1‐mediated H3R17me2a to promote ccRCC progression. Data are shown as mean ± SD. *** p < 0.001. ns means no significance.

Article Snippet: The antibodies used in this study are as follows: LEDGF (Abcam, ab177159), CARM1 (CST, #3379), H3R17me2a (Acive motif, #39 710), PRMT6 (Abcam, ab271091), H3K36me3 (CST, #4909), PPAT (Proteintech, #15401‐1‐AP), PAICS (Proteintech, #12967‐1‐AP), GART (Proteintech, #13659‐1‐AP), ADSL (Proteintech, #15264‐1‐AP), ADSS2 (Proteintech, #16373‐1‐AP), Histone H3 (Proteintech, #17168‐1‐AP), Alpha Actin (Proteintech, #23660‐1‐AP), Flag (Abmart, #M20008), Goat Anti‐Rabbit IgG (H + L) (Proteintech,#SA00001‐2), and Goat Anti‐Mouse IgG (H + L) (Proteintech, #SA00001‐1).

Techniques: Knock-Out, Quantitative RT-PCR, Expressing

Journal: EMBO Reports

Article Title: The L27 domain of MPP7 enhances TAZ-YY1 cooperation to renew muscle stem cells

doi: 10.1038/s44319-024-00305-4

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Rabbit anti-CARM1 , Bethyl , Catalog #IHC-00045.

Techniques: Recombinant, Modification, Magnetic Beads, Sequencing, Blocking Assay, Plasmid Preparation, Electron Microscopy, Software, Reporter Assay, Imaging, In Situ

Journal: iScience

Article Title: Cooperation between PRMT1 and PRMT6 drives lung cancer health disparities among Black/African American men

doi: 10.1016/j.isci.2024.108858

Figure Lengend Snippet:

Article Snippet: PRMT4/CARM1 (C31G9) Rabbit mAb #3379 , Cell Signaling Technology , Cat# 3379; RRID: AB_2068433.

Techniques: Purification, Transduction, Recombinant, Virus, Control, Polymer, SYBR Green Assay, Viability Assay, Mass Spectrometry, Sequencing, Synthesized, Plasmid Preparation, Software, Membrane, Transfection