Fig. 2: CLOCK and BMAL1 interact with RHOA.
From: CLOCK and BMAL1 stabilize and activate RHOA to promote F-actin formation in cancer cells
a Immunofluorescent staining of endogenous BMAL1/CLOCK (red) and RHOA (green) in HeLa and HepG2 cells. Blue color represents the nucleus stained with DAPI. The scale bars represent 10 μm. b The complex was immunoprecipitated from total protein extracts collected from HeLa and HepG2 cells. The immunoprecipitates were analyzed by western blot with antibodies against BMAL1, CLOCK, or RHOA. c Cytosolic (C) and nuclear (N) fractionation followed by Co-IP were performed in HeLa cells. d The TNT® Quick Coupled Transcription/Translation System was used for protein expression in vitro. The synthesized proteins were then analyzed by Co-IP with antibodies against BMAL1, CLOCK, or RHOA. e Two-step IP assay. Cell lysates were immunoprecipitated with the anti-HA antibody, and then BMAL1-HA and the associated proteins were eluted with 4 × HA peptide (30 μmol/μl)
