Fig. 6

Effect of Trolox on mitochondrial function during OB differentiation of hDPSCs. hDPSCs cultured under osteogenic induction medium were continually treated with 500 µM Trolox and OCR (A) and ECAR (B) measured at 7, 14 and 21 days by Seahorse XFe96 Analyzer as in Fig. 3. C OCR/ECAR ratios under basal and maximal activities are shown. Bars values in (A)-(C) are means ± SEM of fold changes of Trolox-treated versus untreated of 3 biological replicates each carried out in triplicate; *P < 0.05; **P < 0.001; ***P < 0.0005. See Fig. 3 for abbreviations. D Energy map obtained plotting the basal OCR and ECAR shown in Fig. 3C, E for Trolox-untreated OB differentiating hDPSCs (white squares) and in panels A and B for Trolox-treated cells (red squares). The values shown are absolute activities and the features on each symbol indicate the differentiating time-points. E Representative cropped immunoblot of the OxPhos complexes protein expression in untreated and Trolox-tretaed cells using a cocktail of specific antibodies as detailed in Methods section. Full-length blots are presented in Additional file 1: Fig. S6. Histogram on the right shows the ß-actin-normalized densitometric analysis as fold changes of Trolox-treated versus untreated hDPSCs; bar values are means ± SEM of data resulting from three independent blots; **P < 0.001 versus untreated