Effect of PGC-1α silencing on RPE mitochondrial replication genes and glycolytic function. Effect of PGC-1α silencing on RPE mitochondrial replication.

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Effect of PGC-1α silencing on RPE mitochondrial replication genes and glycolytic function. Effect of PGC-1α silencing on RPE mitochondrial replication genes and glycolytic function. (A) Live cell imaging of FACS sorted GFP+ shPGC1A and shCtrl ARPE-19 cells confirming the homogeneity of the cell lines. (B) Longitudinal gene expression analysis of the mitochondrial DNA polymerase γ, POLG, and the mitochondrial transcription factor A, TFAM. (C) Glycolytic function measured by ECAR of shCtrl and shPGC1A cells matured for 14 d, after sequential injections of glucose (Glu), oligomycin (Oligo), and 2-deoxyglucose (2-DG). (D) ECAR profile of shCtrl and shPGC1A cells at days 7, 14, and 21 of maturation showing basal ECAR levels (Basal), glycolytic rate (Gly), maximum glycolytic capacity (Gly Cap), glycolytic reserve (Gly Res), and nonglycolytic acidification (Non Gly) (n = 6–12). (E) Quantification of cytoplasmic ATP levels in shCtrl and shPGC1A cells at days 7, 14, and 21 (n = 4). Error bars are means ± SEM. Data were analyzed by multiple unpaired t-test comparisons using the Holm–Sidak method. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001 compared with their respective shCtrl at each time points. Mariana Aparecida Brunini Rosales et al. LSA 2019;2:e201800212 © 2019 Rosales et al.