Ab150376

Total protein was extracted from cells using RIPA Lysis Buffer (Phygene, China) and quantified using the BCA Protein Assay Kit (Phygene). Proteins were separated using 12% SDS-PAGE and transferred to PVDF membranes. The membranes were blocked with 5% bovine serum albumin for 2 h at room temperature and incubated overnight at 4°C with primary antibodies against LRRC15 (1:1,000, ab150376; Abcam, UK), RUNX1 (1:1,000, PA5-85543; Thermo Fisher Scientific, USA), CBF-β (1:1,000, ab231345; Abcam), and GAPDH (1:2,000, #5174; Cell Signaling Technologies, USA). After that, the membranes were further incubated with horseradish peroxidase (HRP)-coupled secondary IgG antibody (1:10,000, ab6721; Abcam) for 120 min at room temperature. Protein bands were examined using an ECL substrate kit (Abcam), and all proteins were quantified based on the internal reference band GAPDH using ImageJ software.