Fig. 7. RhoA lies downstream of N-cadherin in the regulation Igf2 expression. (A) Northern blot analysis (upper panels) of Igf2 mRNA levels in C2 cells transfected with either empty vector (pcDNA), L63 RhoA or T19N RhoA; total RNA is shown by ethidium bromide staining of 18S rRNA. Igf2 P3 promoter luciferase reporter activity (lower panels), in cells co-transfected as above with L63 RhoA, T19N RhoA or control vector (pcDNA), and the Igf2 P3 promoter-reporter construct. Fold activation levels are expressed as the ratio of luciferase activity of L63 and T19N RhoA transfected cells to that in cells transfected with pcDNA: values were normalised to the relative GFP expression levels per unit of protein; ^*P<0.05, ^***P<0.001 (compared with an activation value of 1.0). Western immunoblotting to detect ß-actin is shown as a loading control. (B) Northern blot analysis of Igf2 mRNA levels (upper panels) and Igf2 P3 promoter-reporter activity (lower panels) in C2 cells transfected with cN390, pcDNA or co-transfected with cN390 and L63 RhoA, and examined exactly as for A.