Figure 5: MICU1 negatively regulates OxPhos function and induces lactate production.

(a,b) Oxygen consumption rate (OCR) was measured using the Seahorse XF-96 analyser in OSE (a) and FTE188 cells expressing EV (empty vector) or FLAG-MICU1. An XF trace representing mean±s.e.m. of at least three technical replicates per group is shown. Cells were sequentially treated with 2 μM Oligomycin (O), 0.5 μM FCCP (F) and 1 μM each of Antimycin+Rotenone (A+R) at indicated time points. (c) FCCP stimulated response were derived from the XF trace as in a,b from two independent experiments and represented as mean±s.e.m. (d) CP20 and (e) OV90 cells were transfected with scrambled siRNA (siCTL) or MICU1 siRNA and the oxygen consumption rate (OCR) measured using the Seahorse XF analyser. An XF trace representing mean±s.e.m. from at least three technical replicates per group is shown. (f) FCCP stimulated response were derived from the XF trace of two independent experiments and represented as mean±s.e.m. (g,h) Intracellular lactate was measured in MICU1 silenced OV90 (g) and CP20 (h) cells with treatment with cisplatin 10 μM or vehicle and values represent mean±s.d. (i) Intracellular lactate measurement in MICU1 overexpressing OSE and FTE188 cells as in g. *P<0.05 statistically significant when compared to EV (c,i) siCTL (f–h). Comparisons between two groups were evaluated using two-sided Student’s t-test with unequal variances. Horizontal bar represents statistical comparison between respective groups. All the experiments were repeated independently at least three times and in triplicates.