Fig. 6
From: Akt-mediated phosphorylation controls the activity of the Y-box protein MSY3 in skeletal muscle
Deletion of CSD alters MSY3 nuclear/cytoplasmatic trafficking in myogenic cells. a Graphic illustration of the deleted MSY3 protein domains tested in the mutagenesis assay and distribution of the Akt phosphorylation consensus sites in the MSY3 protein. b Western blot with anti-FLAG Ab of protein extract of C2C12 cells transiently transfected with a FLAG-tagged MSY3 protein (FLAGMSY3) and FLAG-tagged MSY3 protein deleted of the cold shock (FLAGΔCSD), splicing alternative (FLAGΔSHORT), and the carboxy (FLAGΔRP-CD) domains. c FLAG expression tested by IF with an anti-FLAG Ab in C2C12 myoblasts transiently transfected with FLAGMSY3, FLAGΔCSD, FLAGΔSHORT, and FLAGΔRP-CD proteins. The white arrow indicates a nuclear FLAG signal of the FLAGΔRP-CD protein. Scale bar = 100 μm. d N/C ratios of FLAG signal in C2C12 transfected with FLAGMSY3, FLAGΔCSD, FLAGΔSHORT, FLAGΔRP-CD, FLAGSer126Ala, and FLAGSer328Ala proteins, evaluated as a mean ± DS for 60–80 cells from a total of three independent experiments. Quantification was performed as described in Methods. e Akt phosphorylation consensus sites in the MSY3 protein