Anti erk1 2

Whole-cell protein extracts were isolated from primed hPSCs, hEPS, mES and mEPS cells using RIPA lysis buffer (Beyotime Technology Technology, P0013B) supplemented with protease inhibitor cocktail (Thermo Fisher Scientific, 78443) and phosphatase inhibitor cocktail (Thermo Fisher Scientific, 78428). Blots were incubated in 2% BSA/TBST at room temperature for 1 hr, and then, they were incubated with the following antibodies in 5% BSA or 5% skimmed milk powder/TBST at 4°C overnight. For detecting MAPK pathways, the same antibodies were used for human and mouse cells: anti-ERK1/2 (1:1,000; Beyotime Technology, AM076), anti-p-ERK1/2 (1:1,000; Beyotime Technology, AM071), anti-JNK (1:1,000; Beyotime Technology, AJ518), anti-p-JNK (1:1,000; Beyotime Technology, AM516), anti-p38 (1:1,000; Cell Signaling Technology, 9212), anti-p-p38 (1:1,000; Cell Signaling Technology, 9215) and anti-β-ACTIN (1:2,000; Sigma-Aldrich, A1978). For mouse cells, the anti-PARP1 (1:1,000; Santa Cruz, sc-7150) antibody was used. Secondary antibodies were anti-rabbit IgG, HRP-linked antibody (1:5,000; Cell Signaling Technology, 7074) and anti-mouse IgG, HRP-linked antibody (1:5,000; Cell Signaling Technology, 7076), which were incubated 1 hr at room temperature while shaking. The blots were developed using BeyoECL Plus (Beyotime Technology, P0018A).