Abnormal Ribosome Biogenesis Partly Induced p53-Dependent Aortic Medial Smooth Muscle Cell Apoptosis and Oxidative Stress

Figure 3

BOP1 knockdown impaired HASMC protein synthesis rate and motility. (a) After being transfected with scramble (scr) or BOP1 siRNA (si-BOP1) for 48 h, the expression of BOP1, α-SMA, and MLC was detected by western blotting. The expression levels were detected by statistical analysis and shown. (b) Real-time PCR was performed to detect the mRNA level of BOP1, α-SMA, and MLC in HASMCs after being transfected with si-BOP1 or scr for 48 h. (c) HASMCs were transfected with si-BOP or scr for 48 h and administrated with puromycin (1 μg/ml) for 40 min to label the nascent protein. The equal amount of protein was electrophoresed and stained with Coomassie blue to indicate the total protein (left panel). Western blotting was performed to detect the nascent protein by using antipuromycin antibody (middle panel). The statistical analysis of nascent protein/total protein is shown (right panel). (d) Wound healing assay detected the mobility of HASMCs after being transfected with si-BOP1 for 48 h and photographed at the indicated time. The red dotted lines indicated the extent of scratches. (e) The extent of scratches was measured and detected by statistical analysis. Data are representative of three independent experiments and presented as . , , determined by Student’s -test.

(a)

(b)

(c)

(d)

(e)