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Fig. 2. Dominant-negative N-cadherin delays myogenesis and decreases p38 MAP kinase activation and Igf2 expression. (A) Schematic of the dominant-negative cN390 N-cadherin construct. (B) H and E staining of differentiating C2 cells retrovirally transduced with cN390 or pBabe only. Magnification, x50. (C) Western blot of MHC in cN390 or pBabe stably transfected cells; total p38 is shown as a loading control. (D) Western blot to detect the HA tag (cN390 cells only), phospho-p38 (pp38), total p38, phospho-ERK1/2 and total ERK1/2 in vector alone (pcDNA) and cN390 transiently transfected cells. (E) p38 MAPK kinase activity and total p38 levels by western blot analysis in vector alone (pcDNA) and cN390 transiently transfected cells; MBP, myelin basic protein. (F) Northern blot analysis of Igf2 expression in C2 cells stably transfected with either cN390 or pBabe alone; 18S rRNA is shown as a loading control. (G) Igf2 P3 promoter luciferase reporter activity in C2 cells transiently transfected with either cN390 or pcDNA alone. Fold activation levels are expressed as the ratio of luciferase activity of cN390 transfected cells to that of cells transfected with vector alone: values were normalised to the relative GFP expression levels per unit of protein; ^*P<0.05 compared with an activation value of 1.0.

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