Fig. 6

POLRMT pausing impairs renal proximal tubule function. A Confocal image showing RHPS4 (green) uptake into mitochondria following 4 and 24 h of drug treatment. Mitochondria are stained with Mitotracker (grey). Cells in the upper panels are not stained with MitoTracker. Scale bar is 10 µm. B Mitochondrial gene expression in renal proximal tubule epithelial cells (RPTEC) 24 h after RHPS4 treatment, normalized to vehicle treated cells. Gene expression is in arbitrary units (n = 3; P < 0.0001, one-way ANOVA, error bars = S.E.M.). C mtDNA copies per nuclear genome (n = 3; P > 0.05, t-test) (D) mtDNA copies per total DNA mass (n = 3; P > 0.05, t-test). E Oxygen consumption (OCR) is lower (n = 5; P < 0.0001, two-way ANOVA, error bars = S.D.) and (F) Extracellular acidification rate (ECAR) is higher (n = 5, P < 0.0001, two-way ANOVA, error bars = S.D.) in RPTEC 24 h after treatment with RHPS4. G Protein expression of nuclear-encoded TOMM20, mitochondria-encoded COX1 (MT-CO1) and AMPK phosphorylation before and after 72 h of RHPS4 treatment. H Schematic of renal proximal tubule culture system, where cells are seeded on the bottom of the permeable membrane. I Active transport of glucose analogue 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)−2-Deoxyglucose (2-NBDG), expressed as a percentage of transport performed by DMSO-treated cells (n ≥ 5; ***P < 0.001, t-test, error bars = S.E.M)