Skip to main content
. Author manuscript; available in PMC: 2014 Feb 28.
Published in final edited form as: Circ Res. 2013 Sep 30;113(11):1253–1264. doi: 10.1161/CIRCRESAHA.113.301787

Figure 4. Nox4 activation in the ER promotes autophagy in energy-deprived CMs through activation of the PERK/eIF-2α/ATF4 pathway.

Figure 4

Figure 4

A–B. CMs were transduced with Ad-LacZ or Ad-ER-catalase for 48 hours, and then CMs were cultured with normal or glucose-free medium for 4 hours. LC3II levels were evaluated. Representative immunoblots (A) and densitometric quantification (B) are shown. C–D. CMs were transduced with Sh-CT or Sh-Nox4 for 96 hours, and then CMs were cultured with normal or glucosefree medium. Phospho-PERK (Thr980), phospho-eIF-2α (Ser51) and ATF4 protein levels were evaluated. Representative immunoblots (C) and densitometric quantification (D) are shown. N=4. # p<0.05 vs. Sh-CT; * p<0.05 vs. Sh-CT and Sh-Nox4 GD. E. CMs were transduced with Sh-CT or Sh-Nox4 for 96 hours, with and without Sh-PHD4. CMs were cultured with normal or glucose-free medium. ATF4 and LC3 levels were evaluated. F. CMs were transduced with Sh-CT or Sh-Nox4 for 96 hours, together with Ad-LacZ or Ad-PERK for the last 48 hours. CMs were cultured with normal or glucose-free medium. LC3II and phospho-eIF-2α levels were evaluated. G–H. CMs were transduced with Sh-CT or Sh-Nox4 for 96 hours, together with Ad-LacZ or Ad-PERK and with Ad-mRFP-GFP-LC3 for the last 48 hours. CMs were cultured with glucose-free medium. Representative images of mRFP and GFP dots are shown (G), together with quantification of autophagosomes and autolysosomes (H). N=4. Bar=10 μm.