Fig. 7

Comparison of the expression of ion transport regulator IRBIT in Ncbe wt and Ncbe ko mouse CP. a Double immunofluorescence micrograph stained for Ncbe (red) and IRBIT (green) at high magnification of the CP from a Ncbe wt mouse. b The same micrograph showing only the anti-IRBIT immunoreactivity. c A similar micrograph of IRBIT staining in the CP from an Ncbe ko mouse. The fluorescence images are overlaid onto the corresponding DIC images. Arrows indicate the luminal plasma membrane, while arrowheads indicate the basolateral membrane labyrinth. d Immunoblot analysis of the IRBIT abundance in the CP from Ncbe wt and Ncbe ko mice. e Scatter plot comparing relative changes in IRBIT abundance obtained by immunofluorescence microscopy (IF), proteomic mass spectrometry analysis (MS), and immunoblotting (IB)(*p < 0.05, n = 5). Mean values are normalized to control (Ncbe wt) and indicated by horizontal bars. Triangles indicate data points from Ncbe wt CP, whereas circles represent data from Ncbe ko CP. f, g Representative images resulting from PLA assays using anti-Ncbe and anti-IRBIT antibodies on Ncbe wt CP. h, i Similar representative images using the same antibodies on Ncbe ko CP (run in parallel). Positive reaction products are shown in red. Nuclei are shown in blue and the fluorescence signals are overlaid onto the corresponding DIC image