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. 2015 Mar 24;112(14):4447–4452. doi: 10.1073/pnas.1420363112

Fig. 4.

Fig. 4.

Myocd ablation triggers SMC autonomous autophagy and ER stress. (A–G) Myocd gene ablation triggers autophagy in primary cultures of aortic SMCs. MyocdF/F SMCs were transduced with Ad-Cre or Ad-LacZ (control). At 48 h posttransduction, SMCs were fixed and immunostained with antibodies that recognize Cre recombinase or ATG5, ATG7, or LC3. In cells undergoing Cre-mediated Myocd gene deletion (red stain, A–C), robust expression of ATG5, ATG7, and LC3 is observed (green stain, D–F). Original magnification, 400×. (G) Western blot analysis of protein lysates harvested from Ad-LacZ– (Con) or Ad-Cre– (Mut) transduced SMCs incubated with antibodies that recognize Cre, myocardin (Myocd), Beclin-1, ATG-5, ATG-7, LC3, and tubulin (control). Molecular weight markers are shown to the right of the blot. (H–Q) MyocdF/F SMCs were transduced with Ad-LacZ (H–L) or Ad-Cre (M–Q). At 48 h posttransduction, SMCs were fixed and immunostained with antibodies that recognize Cre recombinase or PERK, eIF-2a, ATF4, CHOP, or XBP1. In Ad-Cre–transduced cells, robust expression (green stain) of PERK, eIF-2a, nuclear ATF4, CHOP, and XBP-1 is observed.