Fig. 3

Upregulated cytosolic carbonic anhydrases in TECs. a MCT1 mRNA expression in TECs and NECs was evaluated by RT-qPCR. b MCT1 protein levels in NECs and TECs analyzed by western blotting. c, d, e, f mRNA expression of the pH regulators CAIV, CAIX, CAII, and CAIII evaluated by RT-qPCR in TECs and NECs. g Cellular levels of CAII in NECs and TECs were confirmed by ELISA. h Double immunofluorescence staining for CD31 (red) and CAII (green) in normal mouse tissue (kidney) and A375-SM tumor xenografts. White arrows point to CAII-negative glomerulus and yellow arrowheads to CAII-positive tubules. White arrowheads indicate CAII-positive blood vessels in the A375-SM tumor. Merged image shows the localization of CAII (green) and CD31 (red), and DAPI (blue) Scale bar, 50 μm. i CAII expression in CD31-positive blood vessels in human RCCs (arrows) and absence in normal renal tissue blood vessels (arrowheads). Scale bar, 50 μm. Results presented as mean ± SD, **P < 0.001, by two-tailed unpaired Student’s t-test; N.S., not significant, n = 3)