Figure 4.
Mob2 knockdown increases FlnA phosphorylation at Ser2152 in vitro. (A) C2C12 cells were transiently transfected with EGFP/empty vector control or miRNAs targeting Mob2 (left panel). At 24 h after transfection, cell lysates were prepared and subjected to Western blot analysis using the indicated antibodies. Right panel shows results for the same experiment but performed on age-matched control or patient 1203 fibroblasts. (B) Quantifications representing three biological replicates of (A) summarizing the proportion of phosphorylated Flna at residue 2152 relative to total FLNA and normalized against the loading control Gapdh in C2C12 or age-matched fibroblast controls. Results represented as mean (± SEM). (C) Mean (± SEM) FlnA expression in C2C12 cells expressing control or Mob2 targeting hairpins at the indicated amounts. Expression was normalized against the control using the delta-delta Ct method, as determined by real-time PCR. Expression was normalized against Gapdh and Dimt1 housekeeping genes in the same sample using the relative standard curve method. Mann-Whitney U test; *P < 0.05, **P < 0.005, ***P < 0.001.