Fig. 1

Alteration of mitochondrial proteins and function in NDUFS3-silenced YAPC and MIA PaCa-2 cells. (A-B) YAPC and (C-D) MIA PaCa-2 cells were treated with control RNA (Scrambled, Scr) or with two different NDUFS3 siRNAs (indicated as #1 and #2) and the expression of mitochondrial proteins was evaluated by Western blotting. The efficiency of RNA interference (RNAi) was verified using an antibody against NDUFS3 and HSP90, as loading control. The relative abundance of SDHB and NDUFB8 protein levels was determined through densitometric analysis using Image Lab Software version 6.0.1 normalizing against HSP90 and was reported as the ratio of NDUFS3-silenced cells (#1 and #2) compared to their control (Scr). (E) Mitochondrial function was analyzed in NDUFS3-silenced (#1 and #2) and control (Scr) YAPC cells with the Seahorse Mito stress kit assay. Oxygen consumption rate (OCR) was measured by Agilent Seahorse XF HS Mini analyzer. Basal respiration (F), ATP-production coupled respiration (G), proton leak (H), maximal respiration (I) and non-mitochondrial oxygen consumption (J) were determined by data elaboration with Agilent Seahorse Analytics software. (K) Mitochondrial function was analyzed in NDUFS3-silenced (#1 and #2) and control (Scr) MIA PaCa-2 cells with the Seahorse Mito stress kit assay. Oxygen consumption rate (OCR) was measured by Agilent Seahorse XF HS Mini analyzer. Basal respiration (L), ATP-production coupled respiration (M), maximal respiration (N) and non-mitochondrial oxygen consumption (O) were determined by data elaboration with Agilent Seahorse Analytics software. Values are the mean ± SEM of at least three independent experiments. *p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001