Figure 5

miR-125a-3p and miR-483-5p synergistically regulate the RhoA/ROCK1/ERK1/2 pathway.

(A) hADSCs were transfected with the miR-125a-3p mimic, inhibitor and controls for 72 h. Total protein was extracted for immunoblotting of total ERK1/2 (T-EKR1/2) and phosphorylated ERK1/2 (T-p-EKR1/2). (B) hADSCs were transfected with the miR-125a-3p mimic, inhibitor and controls for 72 h. Nuclear protein was extracted for immunoblotting of n-T-ERK1/2 and n-p-ERK1/2. (C) hADSCs were transfected with the miR-125a-3p inhibitor, miR-483-5p mimic, miR-125a-3p inhibitor plus miR-483-5p mimic, or controls for 72 h. Total protein was extracted for immunoblotting of T-EKR1/2 and T-p-EKR1/2. (D) hADSCs were transfected with the miR-125a-3p inhibitor, miR-483-5p mimic, miR-125a-3p inhibitor plus miR-483-5p mimic, or controls for 72 h. Nuclear protein was extracted for immunoblotting of n-T-ERK1/2 and n-p-ERK1/2. β-actin and PCNA served as loading controls. (E) hADSCs were transfected with miR-125a-3p agomir, pCEP4L-ERK1, miR-125a-3p agomir plus pCEP4L-ERK1, agomir-NC or pCEP4L-NC and induced for differentiation for 12days. Lipid droplet accumulation stained by Oil red O was observed under a microscope. (F) The hADSCs were induced differentiation for 12 days, the protein levels of PPARγ and C/EBPα were detected by western blot. β-Actin served as a loading control.