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Fig. 2 | Skeletal Muscle

Fig. 2

From: Syntaxin 4 regulates the surface localization of a promyogenic receptor Cdo thereby promoting myogenic differentiation

Fig. 2

Overexpression or knockdown of Stx4 promotes or blocks myoblast differentiation, respectively. a Lysates of control or Stx4-overexpressing C2C12 cells were immunoblotted with antibodies against Stx4 and pan-Cadherin as a loading control. The relative signal intensity of Stx4 to pan-Cadherin was quantified and added under the blot. b Lysates of control or Stx4 expression vector transfected C2C12 cells from the differentiation day 1 (D1) and D2 were immunoblotted with antibodies to MHC, MyoD, Myogenin, Troponin T, and pan-Cadherin as a loading control. c Control or Stx4 expression vector transfected C2C12 cells were induced to differentiate for 3 days and immunostained with MHC antibody followed by DAPI staining to visualize nuclei. Size bar = 100 μm. d The quantification of myotube formation shown in panel c. Values represent means of triplicate determinations ± 1 SD. The experiment was repeated three times with similar results. Significant difference from control, *p < 0.01. e Control or shStx4 expression vector transfected C2C12 cells were analyzed by immunoblotting with antibodies to Stx4 and pan-Cadherin as a loading control. The relative knockdown levels of Stx4 to pan-Cadherin are quantified and added under the blot. f Control or shStx4 expression vector transfected C2C12 cells were induced to differentiate for 2 or 3 days and then lysates were subjected to immunoblotting with antibodies to MHC, MyoD, Myogenin, Troponin T, and pan-Cadherin as a loading control. g Control pSuper or shStx4 expression vector transfected C2C12 cells were induced to differentiate for 3 days and immunostained with an antibody to MHC followed by DAPI staining to visualize nuclei. Size bar = 100 μm. h The quantification of myotube formation shown in panel g. Values represent means of triplicate determinations ± 1 SD. The experiment was repeated three times with similar results. Significant difference from control, *p < 0.01, **p < 0.005

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