NOX5 and p22phox interact in Mo-DC. (A) Representative western blot of co-IP. p22phox (left) and NOX5 (right) were immunoprecipitated with antibodies to p22phox and NOX5, respectively, and 2 different NOX5 antibodies were used for detection. IP (immunoprecipitate) fractions with or without antibodies (−Ab and +Ab); SUP (supernatant) after immunoprecipitation. (B) Total lysates were separated by 2-dimensional BN/SDS-PAGE (4%-16% and 12%) and immunoblotted with NOX5 and p22phox antibodies. The NOX5-p22phox complex is shown in the red box. (C) Confocal microscopy of Mo-DC, labeled with anti-NOX5 and Alexa 488, anti-p22phox and Alexa 555, and 4′,6-diamidino-2-phenylindole (DAPI; nuclei); 40× objective, zoom 6×. (D) Representative image using the Duolink proximity ligation assay in Mo-DC. Interaction between NOX5 and p22phox is represented by red dot signals (right panel). In the negative control (left, without primary antibodies), only the DAPI signal can be observed; 63× objective, zoom 2×.