Fig. 3.
Effect of Hand2 knockdown on TH expression in cultured sympathetic neurons. E12 sympathetic neurons were transfected with control GFP siRNA (A,B) or Hand2 siRNA (C,D) and analyzed after 2 days in culture for the expression of TH mRNA expression by in situ hybridization. A sense RNA probe was used to control for the specificity of the TH in situ hybridization signal (E,F). TH-positive neurons display a strong in situ hybridization signal (B,D), which is completely absent in non-neuronal cells and in neurons treated with the sense control RNA (F). In cultures treated with Hand2 siRNA the proportion of TH-expressing sympathetic neurons decreased from 80% in control transfections to about 40% in cultures transfected with Hand2 siRNA. (A,C,E) Phase contrast; (B,D,F) bright field optics. Data shown in (G) represent mean ± s.e.m. (n=3), **P<0,01; ***P<0,001.