Fig. 5.
ATG7 ablation results in accumulation of aberrant mitochondria, oxidative stress, and activation of protective and counteractive responses. (A) EM images showing accumulation of abnormally looking mitochondria (arrow) in 12-wk-old mice. [Scale bars, 2 μm (Left), 250 nm (Top Right), and 500 nm (Bottom Right).] Healthy vs. damaged mitochondria were quantified by stereological analysis. Results are means ± SEM n = 7–8 micrographs per condition (one mouse for each condition). ***P < 0.001. (B) Mfn1 and Parkin expression in pancreata of above mice. Protein amounts were determined by densitometry and normalized to actin (Bottom). (C) Protein expression of NRF2 and NQO1 in primary acinar cell (NRF2), and total prancreata samples (NQO1) of 12-wk-old Atg7F/F and Atg7Δpan mice. (D) Expression of the indicated pancreatic mRNAs was determined by qPCR. Results are means ± SEM of triplicates. n = 3 mice per condition. *P < 0.05, **P < 0.01 vs. Atg7F/F. (E) IB analysis of antiapoptotic proteins. (F) IB analysis of total and phosphorylated AMPKα in 12-wk-old pancreata. (G) IB analysis of phosphorylated mTORC1 targets in pancreatic lysates. (H) Protein synthesis in primary acinar cells of Atg7∆pan and Atg7F/F mice, quantified as the ratio of [3H]-leucine incorporated into TCA insoluble material between control and cycloheximide (30 µg/mL) treated cells. *P < 0.05 vs. Atg7F/F. Results are means ± SEM of triplicates. n = 3 mice per condition.