Extended Data Fig. 6: Loss of ciliogenesis impairs flow-induced mitochondrial biogenesis in Ift88-knockout KECs.
(a, b) WT and Ift88-/- KECs were subjected to flow, or not, for 4 days, and Ppargc1a and Tim23 mRNA levels were quantified by RT–qPCR, normalized to β-actin, and presented as fold increases. Data in a, b) are means ± SEM, n = 3 independents experiments. The statistical significance of a, b) was calculated by a two-tailed t-test. (c–e) Wild-type (WT) and Ift88-/- KECs were subjected to flow, or not, for 4 days, and levels of mitochondrial proteins TOM20, TIM23, and were visualized by western blot analysis. Representative blot from n = 3 independent experiments. (d, e) The TOM20/Actin and TIM23/Actin ratios were determined by densitometry. Data are means ± SEM, n = 3 independents experiments. The statistical significance of d) and e) was calculated by a two-tailed t-test. (f, g) Images of WT and Ift88-/- KECs subjected to flow for 1 day, or not, fixed, and labeled with anti-LC3 antibody (green), LipidTox to stain LDs (red), and Hoechst 33342 to stain nuclei (blue). Scale bars 5 μm. Representative images from n = 3 independent experiments. (g) Relative LC3-LD co-localization was quantified by Pearson’s coefficient analysis. Data are means ± SEM. Individual data points correspond to single images analyzed from n = 3 independent experiments. The statistical significance was calculated by a two-tailed t-test. Data and unprocessed blots are available as source data.
