Figure 4From: Differential regulation of p53 function by the N-terminal ΔNp53 and Δ113p53 isoforms in zebrafish embryosZebrafish survival in response to simultaneous ectopic expression of all three p53 isoforms in zebrafish embryos. (A) Capped mRNAs encoding FLp53 (0.1 ng mRNA/embryo) alone or in combination with ΔNp53 (1 ng mRNA/embryo) or Δ113p53 (1 ng mRNA/embryo) or ΔNp53 (1 ng mRNA/embryo) together with Δ113p53 (1 ng mRNA/embryo) were injected into 1-4 cell embryos and survival was scored over seven days. Standard error was determined using triplicate dishes of 60 embryos each. Capped mRNA of green fluorescent protein (GFP; 2 ng RNA/embryo) was used as a negative control. (B) Molecular interaction of zebrafish p53 isoforms as determined by coimmunoprecipitation experiments using Saos-2 cells. All immunoprecipitations were performed using the HA Tag IP/Co-IP kit (Pierce, Rockford IL). The immunoprecipitates were subjected to SDS-PAGE and immunoblotted using antibodies recognizing the HA- and Myc-tags. The upper panel shows immunoblot detection of the HA-tag whereas the lower panel shows blots probed with antibodies detecting the Myc-tag. Experimental conditions were as follows: (1) Transfection control, (2) HA-Akt and Myc-ΔNp53 (3) HA-FLp53 and Myc-ΔNp53, (4) HA-FLp53 and Myc-Δ113p53, (5) HA-Δ113p53 and Myc-ΔNp53. The asterisk denotes the HA-Δ113p53 band. Expression of transgenes was validated by Western blot analysis of whole cell extracts (see Additional file 2).Back to article page