Anti prmt2

The paraffin-embedded tissue sections were used for examination of HE staining. For the IHC staining, specimen’s tissue slides were de-paraffinized, rehydrated, and antigen retrieval. The tissue slides were blocked for at least 1 h at room temperature and then incubated with appropriate primary antibodies overnight at 4 ℃. The antibodies included anti-PRMT2 (1:100; Abcam), anti-CCNB1 (1:200; Santa Cruz Biotechnology), anti-CCND1 (1:100; Absin), anti-CDK4 (1:200; Wanleibio), and anti-H3R8me2a (1:500; Novus Biologicals). After careful washing, the slides were incubated with horseradish peroxidase (HRP) conjugates using DAB detection. For the IHC analysis, we quantitatively scored the tissue sections according to the percentage of positive cells and staining intensity. We assigned the following proportion scores: 1 if 0–25% of the tumor cells showed positive staining, 2 if 26–50% of cells were stained, 3 if 51–75% stained, and 4 if 76–100% stained.

Cultured cells were fixed with 4% formaldehyde for 5 min, permeabilized with 0.1% Triton X-100 in phosphate-buffered saline (PBS) for 10 min and then incubated with 5 mg/ml bovine serum albumin for 60 min at room temperature. Immunostaining was performed using the appropriate primary antibodies overnight at 4 ℃: anti-PRMT2 (1:100; Abcam) and anti-H3R8me2a (1:500; Novus Biologicals). After careful washing, the cells were incubated with FITC 490 secondary antibody (ZSBIO, dilution 1:200) for 1 h at room temperature. Nuclei were counterstained with Hoechst.