Fig 2. SERCA1b silencing modifies the proteins responsible for differentiation.
Effect of decreased SERCA1b protein level on the expression pattern of key proteins involved in skeletal muscle differentiation. (A-C) Western-blot analysis to detect the main differentiation marker proteins (MyoD, and Calcineurin) on the 5th day of differentiation. Total protein samples were used (30 μg in each lane) to examine the protein expression level. Representative data each of 3 independent experiments. Actin was used as a control. (D-E) mRNA expression pattern of myostatin and MCIP 1.4 was assessed by RT-PCR reaction using specific primers and detected at the expected size. GAPDH was used as a control. (F) Activity of calcineurin was assayed by using RII phosphopeptide substrate, and the release of free PO4 3- was detected by the malachite green assay. Six separate wells were used from every experimental group. Measurements were carried out in two independent experiments. Asterisks (*) mark significant (P<0.05) differences between cells.