Fig. 5

MUC13-Glut-1 interaction is disrupted by Sulfasalazine. a Confocal immunofluorescence of MUC13 and Glut-1 in MUC13-expressing, HPAF-II, and AsPC1 cells depicting co-localization (yellow color). Cells were cultured in 4-well chambered slides, fixed, permeabilized, stained with indicated antibodies and analyzed for confocal microscopy. DAPI was used as a counter stain for the nucleus. b Cells were treated with sulphasalazine (0.9 mM) for 24 h and processed for immunostaining and confocal microscopy. Images were captured at ×400. c, d Reciprocal co-immunoprecipitation assay between MUC13 and Glut-1 performed using protein lysates from HPAF-II cells. Cells were treated with sulfasalazine (0.9 mM) for 24 h and protein lysates prepared. The immunoprecipitates were resolved on a 10% gel, and followed by immunoblotting with anti-MUC13 and anti-Glut-1 antibodies. e Lactate assay was performed in MUC13-expressing and MUC13-null cells after treatment with or without sulfasalazine. Cell culture media was collected after 48 h to measure the amount of l-lactate concentration using lactate assay kit