FIGURE 7.

CANX and NOX4 interaction in proximity ligation assay analysis. A, representative images of proximity ligation assay analysis showing the interaction of NOX4 with CANX in HEK293 cells with (NOX4) and without (Ctl) overexpression of NOX4. Positive signal of interaction is shown as green dots, nuclei are stained with DAPI (gray). In the two left images the primary antibody against NOX4 was omitted, in the right images, a blocking peptide against calnexin was added. B and E, representative images of proximity ligation assay analysis showing the interaction of NOX4 with CANX in podocytes from wt or Nox4^−/− (N4^−/−) mice (B) or MEF from wt or Canx^−/− mice (E). C and F, dots per nuclei were analyzed of at least three independent experiments (8 pictures each) and normalized to wt cells. CANX antibody blocking peptide was used as control, n ≥ 3, mean ± S.E., *, p < 0.05; **, p < 0.01; ***, p < 0.001 relative to the corresponding knock out; #, p < 0.05, ##, p < 0.01 relative to the control without blocking peptide D. G, representative Western blot characterizing the Nox4-deficient podocytes (D) or Canx-deficient MEFs (E). As loading control β-Tubulin (C, β-Tub.) or β-Actin (G, β-Act.) was used.