Fig. 3

Alterations of mitochondrial and lysosomal biogenesis regulators characterize chemoresistant cells. A, B Relative protein abundance of SOD1, SOD2, PRX, and CAT was assessed by Western blot analysis and quantified by densitometry normalizing against HSP90. C, D Relative protein abundance of PGC-1α, NRF-1, and TFAM was assessed by western blot analysis and quantified by densitometry normalizing against β-Actin. E, F Relative protein abundance of mTOR, mTOR p-2448, Raptor, Rictor, GβL, S6K, p-S6K, Parkin, and PINK1 was assessed by Western blot analysis and quantified by densitometry normalizing against HSP90. G, H Colocalization rates were determined for A2780 and A2780 CIS cells immunostained with anti-mTOR (red) and anti-LAMP-1 (green) antibodies. Nuclei were stained with DAPI (blue). White boxes indicated zoomed areas on the right. Scale bar = 10 μm. I, J The total TFEB abundance was determined by Western Blot analysis and quantified by densitometry normalizing against HSP90 using Image Lab software (Bio-Rad). K, L Relative protein abundance of the nuclear fraction of A2780 and A2780 CIS subjected to Western blotting using anti-TFEB, and anti-Histone H3, used as the loading control, was determined. For densitometry Image Lab software (Bio-Rad) was used. Data represent the mean ± SEM of at least three independent experiments. * p < 0.05; ** p < 0.01; ***p < 0.001