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Figure 1


Fig. 1. Quantification of TRPC channels in control and TRPC1-knockdown myoblasts. (A) Quantification of TRPC channels in C2C12 myoblasts. Each cDNA was amplified in duplicate and Ct values were averaged for each duplicate. The average Ct value for cyclophilin D was subtracted from the average Ct value for the gene of interest, giving a {Delta}Ct. Note that the {Delta}Ct for TRPC1 is at least 7 units below other values, meaning the expression is at least 128 times higher than for any other TRPC. (B) Western blot analysis of TRPC1, MARCKS and β-actin expression in myoblasts transfected with control siRNA (lanes 1 and 3) or with one of two different TRPC1 siRNAs (lanes 2 and 4). TRPC1, MARCKS and β-actin proteins were detected sequentially on the same blot (stripped twice). Blot on left: 12 µg protein/lane; Blot on right: 4 µg/lane. Results are representative of three measurements. (C) Western blot analysis of TRPC1 expression in nontransfected C2C12 cells (lane 1) and in C2C12 cells stably expressing TRPC1 shRNA (lane 2) or control shRNA (lane 3), after 7 days of differentiation. Loading: 10 µg protein/lane. Left panel, Ponceau Red staining; Right panel, immunodetection with TRPC1 antibody.