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. 2015 Sep 22;11(11):2014–2032. doi: 10.1080/15548627.2015.1096485

Figure 5.

Figure 5.

SQSTM1 accumulation links defective VSMC autophagy to senescence. (A) Atg7+/+ VSMCs were transfected with 5 µg plasmid DNA encoding SQSTM1 (SQSTM1+). Four d after transfection, VSMCs were analyzed for SQSTM1, CDKN2A, phospho RB and total RB expression by western blotting. (B,C) SQSTM1 overexpressing VSMCs were incubated with BrdU (B) to examine proliferation capacity (***, P < 0.001; n = 2 counting regions of 1000 cells/region/condition in duplicate; Student t test) or (C) stained with X-gal mixture for 24 h to quantify the number of senescence-associated GLB1-positive VSMCs (***, P < 0.001; n = 3 counting regions of 150 cells/region/condition in duplicate; Student t test). Scale bar: 50 µm.