Dystroglycan Phosphorylation and Ubiquitination

Dasatinib was obtained from Selleckchem (Munich, Germany) and Pyr-41 and MG132 from Sigma-Aldrich (Gillingham, UK). The following antibodies were used in WB and/or immunofluorescence (IF) applications. Non-phosphorylated β-dystroglycan [MANDAG2; WB 1:100, IF 1:100 (47 (link))], β-dystroglycan phosphorylated at tyrosine 892 [1709; WB 1:500 (13 (link),19 (link))], α-tubulin (WB 1:3500, Sigma-Aldrich), ubiquitin (WB 1:100, Enzo Life Sciences), biotinylated Concanavalin A (WB 1:4000, Vector Labs), western blots were detected by species-specific horseradish peroxidase conjugated to secondary antibodies or to streptavidin (1:5000, Sigma-Aldrich) and detected by ECL. In immunofluorescence, primary antibodies were detected by species-specific Alexa Fluor-conjugated secondary antibodies (IF 1:200, Molecular Probes) and F-actin was detected with rhodamine-conjugated phalloidin (IF 1:20, Molecular Probes).