Research Paper Volume 15, Issue 15 pp 7513—7532

Figure 5. NORAD knockdown increased the PGC-1α acetylation and mitochondrial ROS. ARPE-19 cell lines were transfected with siRNA of siNORAD or siNC, after 96 hours, the cells were observed for mitochondrial damage, mitochondrial ROS, PGC-1α. (A) NORAD knockdown decreased the level of mRNA of mitochondrial transcription factor A (TFAM), mitochondrial DNA polymerase catalytic subunit (POLG), mitochondrially encoded NADH dehydrogenase 1 (ND1), and mitochondrially encoded NADH dehydrogenase 5 (ND5) in irradiation treated ARPE-19. (B) Flow cytometry was used to detect the mitochondrial ROS levels. Irradiation increased the mitochondrial ROS and NORAD knockdown aggravates irradiation-induced the rise of mitochondrial ROS. Data were from three separate experiments and described as mean ± SD. **P < 0.01. (C) NORAD knockdown aggravated irradiation-induced ROS production. The ROS levels were observed under an inverted fluorescence microscope. Images were representative of three separate experiments. n = 3, **P < 0.01. (40X, measure scar 20um) (D) Immunoprecipitation and western blot analysis of acetyl-PGC-1α levels in ARPE-19 transfected with siNORAD and exposed to sodium iodate. (E) Immunoprecipitation and western blot analysis of acetyl-PGC-1α levels in ARPE-19 transfected with siNORAD and exposed to irradiation. (F) Immunoprecipitation and western blot analysis of acetyl-PGC-1α levels in heart, liver, lung, kidney, brain, and eye of NORAD knockout mouse.