Figure 3.

Knockdown of NAE1 induces G2/M arrest and apoptosis in hSMCs. (a) QRT-PCR verification of gene-specific silencing of NAE1 in hSMCs. NC siRNA: negative control siRNA. (b) Western blotting analysis of Nedd8-cullins in hSMCs. GAPDH was used as a loading control. (c) FACS analysis of cell cycle of hSMCs. (d) Quantification of the percentages of hSMCs in different cell cycle phases. (e) TUNEL staining of hSMCs. (f) Quantification of TUNEL-positive cells. (g) Western blotting analysis of cleaved caspase 3 in hSMCs. (h) Quantification of (g). Cells were transfected with siRNA for 12 h (a) or 24 h (b–h), and then cultured for another 72 h. Scale bar: 200 μm. Each experiment was repeated at least three times. *P<0.05, **P<0.01, ***P<0.001