Fig. 3

GCN5L1 affects the intracellular distribution of TFAM and then alters TFAM-mtDNA binding capability. A, B mRNA levels of TFAM were detected by qRT-PCR in GCN5L1 overexpression or knockdown TECs. C–F Protein levels of TFAM were detected by western blot in GCN5L1 overexpression or knockdown TECs. G, H Confocal microscopy for the accumulation of TFAM in mitochondria in TECs co-transfected GCN5L1 overexpressing plasmid or GCN5L1 silencing siRNA with pDsRed2-Mito plasmid. I–LWestern blot showed the TFAM expression in isolated mitochondrial protein in GCN5L1 overexpression or knockdown TECs. M–P The binding of TFAM with mtDNA revealed by ChIP assay in GCN5L1 overexpression or knockdown TECs and rabbit IgG was used as a negative control. ns: not significant, *P < 0.05, **P < 0.01, and ***P < 0.001