Immunolocalization and quantification of mitochondrial and EMT markers in PGC-1–floxed mice 4 mo after subretinal AAV injection. Immunolocalization and.

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Immunolocalization and quantification of mitochondrial and EMT markers in PGC-1–floxed mice 4 mo after subretinal AAV injection. Immunolocalization and quantification of mitochondrial and EMT markers in PGC-1–floxed mice 4 mo after subretinal AAV injection. (A) Representative images of RPE flat-mount preparations from AAV-GFP and AAV-Cre–injected animal at 4 mo immunostained for the mitochondrial marker, COXIV, and the EMT markers, Coll. VI, Vimentin, ZEB1, and TWIST1 (red) and costained with DAPI (blue). Whereas depletion of COXIV was observed in all fields of AAV-Cre–injected mice, increases in the EMT markers were only observed in localized areas with abnormal and sloughed RPE (arrows). Insert in the ZEB1 panel shows nuclear localization of the EMT transcription factor in amorphic RPE cells (n = nucleus). Scale bar is 50 μm. (B) Fluorescence intensity quantification indicated a significant reduction of COXIV mean fluorescence intensity by 60% after AAV-Cre. Mean fluorescence intensity of the EMT-markers was not significantly altered as the changes were localized (4–6 randomly selected fields per sample, n = 3). Error bars are means ± SEM. Unpaired t tests were used in all comparisons. **P ≤ 0.01 compared with their respective AAV-GFP group. Mariana Aparecida Brunini Rosales et al. LSA 2019;2:e201800212 © 2019 Rosales et al.