Fig. 2: HSP60 maintains mitochondrial function in PDAC cells.

a, b Analysis of the oxygen consumption rate (OCR) of various cell lines by an Oxygraph-2k comparing shHSP60 Panc-1 cells with a control cells and b rescued expression of HSP60 cells. OCR was first measured of each cell line under basal conditions followed by sequential addition of oligomycin (100 μg/mL) at the indicated ATP-linked OCR. Basal, basal OCR; Oligomycin, OCR in the presence of 100 μg/mL oligomycin; Basal-oligomycin, OXPHOS-associated OCR was calculated by subtracting the OCR in the presence of oligomycin from basal OCR. c Measurements of mitochondrial ATP levels. Cells were incubated with 10 mM glucose or 5 mM 2-DG plus 5 mM pyruvate to determine mitochondrial ATP generation. Relative average ATP levels in mitochondria per cell line are shown. d Respiratory chain supercomplexes and single complexes in shHSP60 Panc-1 cells and control cells. e The respiratory chain supercomplex in shHSP60 Panc-1 cells and cells with rescued expression of HSP60. f Mitochondrial ATP levels and g respiratory chain supercomplexes were measured in cells from PDAC cell lines (SW1990, Patu-8988, and Panc-1) compared with normal pancreatic ductal cells (hTERT-HPNE). h Respiratory chain supercomplexes in fresh PDAC tissue samples compared with those found in paired normal tissues. The values of OCR and ATP generation were normalized to protein concentration. Blots were probed with anti-Grim19, anti-UQCRC2, anti-SDHA or anti-COX1, and anti-cyt c for complex I (CI), complex II (CII), complex III (CIII), and complex IV (CIV), respectively. All data are presented as mean ± SEM (n ≥ 3). *P < 0.05, **P < 0.01, and ***P < 0.001