HSPB1 Regulates Autophagy and Apoptosis in Vascular Smooth Muscle Cells in Arteriosclerosis Obliterans

Figure 2

HSPB1 regulates apoptosis in VSMCs. (a) VSMCs were treated with 0, 15, 30, 45, 60, or 75 g/ml ox-LDL for 24 hr, and then, HSPB1 protein expression was measured by Western blotting. (b) Densitometry was used to calculate the fold change in the expression of HSPB1 relative to the expression of β-actin. (c–h) VSMCs were treated with pCMV-Cmyc-HSPB1 and HSPB1-siRNA for 48 hr, followed by 60 g/ml ox-LDL for 24 hr. (c) Cell lysates were harvested to examine the protein levels of PCNA, Ki67, and β-actin by Western blotting. (d) Densitometry was used to determine the fold change in the expression of PCNA and Ki67 relative to the expression of β-actin ( for each group; , , and ). (e) Representative flow cytometric plots are presented. (f) The data are presented as (). (g) Western blotting was performed as described above, and the protein levels of Bcl-2, Bax, cleaved caspase-3, and β-actin were measured. (h) The levels of these proteins were quantitated by densitometric analysis using ImageJ and normalized to the level of β-actin. The results are presented as the fold changes in expression compared with those in the control group ( for each group; , , and ).

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