Fig. 1: C15ORF48 expression activates AMPK-ULK1 signaling.

a A549 cells stably expressing C15ORF48 (A549/C15ORF48) or the empty (A549/empty) vector were fixed and stained with anti-C15ORF48 antibody and MitoTracker. Nuclei were counter-stained with TO-PRO-3. Representative images are shown from three independent experiments. b A549/empty and A549/C15ORF48 cells were analyzed for mitochondrial membrane potential by staining with TMRM (30 nM) for 30 min. Median TMRM fluorescence intensities are shown as the mean ± SD. Statistical significance was calculated using two-tailed unpaired Student’s t test (n = 3, biological replicates). c A549/empty and A549/C15ORF48 cells were analyzed for intracellular ATP levels. ATP amount in a cell is shown as the mean ± SD. Statistical significance was calculated using two-tailed unpaired Student’s t test (n = 3, biological replicates). d A549/empty and A549/C15ORF48 cells were lysed and subjected to western blotting with indicated antibodies. Band intensity was measured, and quantitative ratios are shown. Data are representative of three independent experiments with three biological replicates.