Fig 1: MPP is important for PyB6-TA cells. For simplification, miR-Es are indicated by sh. (A, B) Analysis of Pmpca (A) and Pmpcb (B) protein expression by Western blot in miR-E-transduced PyB6-TA cells cultured for 4 (D4) or 8 days (D8) ± Dox. Left: Quantification of Western blot data; mean reduction of protein level ± SD relative to uninduced cells normalized to tubulin or GAPDH. Right: Representative Western blots; tubulin or GAPDH as loading control; 25 µg protein loaded. M: marker. (C, D) Plate colony formation assay of miR-E-transduced PyB6-TA cells cultured ± Dox under normoxic conditions (21% O2, 8–9 days) or hypoxic conditions (3% O2, 12–14 days). ShRenilla-transduced cells as control. (C) Exemplary pictures. (D) Average cell growth reduction ± SD relative to untreated. Significance (p) one-sample t-test to 0 or two-sample t-test, equal variances assumed (n = 5–9). (E, F) Flow cytometry-based analysis of mitochondrial activity in miR-E-transduced PyB6-TA cells cultured for 4 or 8 days ± Dox by deep Red FM MitoTracker staining. (E) Exemplary median APC intensity. (F) Median APC fluorescence intensity ± SD relative to uninduced. Significance (p): one-sample t-test to 0 (n = 5–6). Ns, not significant.