Mouse anti setd4

Total cellular proteins were extracted by RIPA lysis buffer (Beyotime P0013B) containing protease inhibitor cocktail (MedChemEx-191 press; HY-K0010); Tissue total proteins were prepared using the Trizol Extraction Reagent (Invitrogen #15596018) according to manufacturer instructions. Protein concentration was resolved by SDS-PAGE and transferred to PVDF membranes (Bio-RAD #1620177). The filters were blocked in 1% BSA for 1 h at room temperature and then incubated with primary antibody overnight at 4 °C. The filters were washed 3 times in TBS and incubated in secondary antibodies overnight. Membrane-bound immune complexes were detected by ultra-sensitive enhanced chemiluminescence system (Bio-RAD #102031152) on Amersham Imager 600 (GE). Primary antibodies were used as below: mouse anti-SETD4 (1:50, Santa Cruz Biotechnology sc-514060); Full antibody details are provided in the supplemental information antibodies list. Quantification was performed by densitometric analysis using ImageJ 64 software.

The following antibodies were used: Rat anti-BrdU (1:100, Abcam, ab6326); Rabbit anti-Ki67 (1:100, Abcam, ab16667); Rabbit anti-H3pS10 (1:150, Epitomics, 1173-1); Mouse anti-Setd4 (1:50, Santa Cruz Biotechnology, sc-514060); Rabbit anti-H4K20me3 (1:200, Abcam, ab9053); Mouse anti-HP1α (1:200, EMD Millipore Corp, 05-689); Rabbit anti-H3K9ac (1:200, Epitomics, 1328-1); Mouse anti-Troponin T (1:100, Invitrogen, MA5-12960); Mouse anti-sarcomerica-actin (1:100, Sigma, A2172); Rat anti-CD31 (1:200, BD Biosciences, 5553370); Goat anti-VE-cadherin (1:200, RD, AF1002); Rabbit anti-PDGFRα (1:100, RD, AF1062); Rabbit anti-Sox9 (1:100, Abcam, ab185230); Rabbit anti-α-SMA (1:150, Sigma, A2547); Rabbit anti-FABP4 (1:150, Abcam, ab92501); Rat anti-c-Kit (1:100, Abcam, ab25022).