Western Blot Analysis of RAB37 Expression

Western blots were performed following our previous protocols 19 (link). 293T cells were transfected with RAB37-FLAG or vector control. At 48 h after transfection, the whole protein extracts were extracted from the cells and used for SDS-PAGE. The gels were transferred onto a 0.45 μm PVDF membrane (Cat# NK0414, Roche Diagnostics, Indianapolis, IN, USA). The membranes were blocked with 5% skim milk powder (Cat# 1172GR100, BioFroxx Gmbh) in TBST and incubated with the primary antibody overnight at 4 °C followed by the horseradish peroxidase-conjugated secondary antibody at room temperature for at least 1 h. ECL Plus detecting reagents (Millipore, Billerica, MA, USA) were used to visualize the protein bands. ImageJ (version J2, NIH, Maryland, US) was used to analyze band intensity values.