Fig. 6
Multiprotein complex is properly assembled in L6 myotubes overexpressing Panx1. Double-staining immunofluorescence was performed using two different primary antibodies in the same sample, as detailed: anti-Strep/anti-Panx1 (a), anti-DHPR/anti-Panx1 (b), and anti-Cav3/anti-Panx1 (c). Bar graphs next to the image panels show Manders’ coefficients for the co-localization analysis. Scale bar = 10 μm. White boxes indicate area that is then magnified in the row below. Bars correspond to the average of the data obtained from three independent experiments, where three regions of interest (ROI) were evaluated per image for the Manders’ coefficient quantification. d Co-immunoprecipitation assay was performed in L6-Panx1 myotubes. Anti-Panx1, anti-DHPR, and anti-P2Y2R antibodies were used to immunoprecipitate whole lysates from L6-Panx1 cells. Panx1, DHPR, and P2Y2R all co-immunoprecipitated in these cells together with Dys and Cav3. Representative blot of the three experiments is shown