Extended Data Fig. 2: Effect of fluid flow on mitochondrial biogenesis and activity in cells.
(a) KECs and IMCD3 were subjected to flow for 4 days. Megalin and Aqp2 mRNA expression was then visualized by real-time RT–qPCR and normalized with respect to β-actin, and presented as fold increases. Data are means ± SEM. n = 3 independents experiments The statistical significance was calculated by a two-tailed t-test (b) KECs and IMCD3 were subjected or not (d0) to flow for 1 to 4 days (d1, d2, d4), and the levels of mitochondrial proteins (Tom20, Tim23, MTCO1) were visualized by western blot analysis. Representative blot from n = 3 independent experiments. (c) Tom20/Actin and Tim23/Actin ratios were determined by densitometry, Data are means ± SEM, n = 3 independents experiments. The statistical significance was calculated by a two-tailed t-test (d) KECs and IMCD3 were subjected or not (d0) to flow for 4 days (d4), and Ppargc1a mRNA expression was visualized by real-time RT–qPCR. Data are means ± SEM, n = 3 independents experiments. The statistical significance was calculated by a two-tailed t-test (e) KECs and IMCD3 were subjected to flow or not (D0) for 4 days (D4), harvested by trypsinization, and mitochondrial DNA was quantified by qPCR analysis. Data are represented as ratios between D4 and D0, Data are means ± SEM, n = 3 independents experiments. The statistical significance was calculated by a two-tailed t-test (f) KECs and IMCD3 were subjected to flow (shear) or not (static) for 4 days and then harvested by trypsinization. The oxygen consumption rate (OCR, Seahorse XF Mito Stress Test) was measured using the Seahorse extracellular flux assay; shown are f) Seahorse trace representative of n = 3 independent experiments and g) a histogram representative of n = 3 independent experiments, data are means ± SEM of 4 biological independent OCR results. The statistical significance was calculated by a two-tailed t-test. Data and unprocessed blots are available as source data.
