Fig. 4.

sihNAA30-treated cells display fragmented mitochondria. HeLa (A, B) and CAL-62 (C, D, E) cells depleted for Naa30 were immunostained for the mitochondrial marker COX IV-mouse (A) or COX IV-rabbit (C, E). Micrographs were taken with a confocal and Z-stacks were Z-projected to visualize COX IV. DAPI (blue) was used to visualize the nuclei (A, C) in overlay with COX-IV (red). The confocal micrograph in (E) was taken with a 6.5 zoom and deconvoluted. White bars correspond to 10 μm. The percentages of cells with perinuclear and fragmented mitochondria were calculated for each sample (B, D). Data are mean and standard deviations from three independent experiments (n = 100–150). The difference between siCTR and sihNAA30 treated cells was statistically significant (p < 0.05, student t test). (F) Immunoblots of cell lysates from HeLa cells treated with nontargeting siRNA (siCTR) or sihNAA30. Cells were harvested 72 h post siRNA transfection. Blots were probed with anti-Naa30 to assess levels of endogenous Naa30. β-tubulin was used as loading control.