Skip to main content


Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Figure 6 | BMC Molecular Biology

Figure 6

From: Regulation of p73 by Hck through kinase-dependent and independent mechanisms

Figure 6

Hck inhibits the transcriptional activity of p73α and β isoforms, dependent on its SH3 domain. (A-D) Hela cells transfected with pCMV-βgal (50 ng) and different promoter constructs (100 ng) along with p73α or p73β and Hck or KD-Hck were subjected to either luciferase or CAT activity measurements. The amount of DNA was kept constant in all transfections to 400 ng by adding pcDNA3. Hck, KD-Hck, mSH3-Hck and c-Abl plasmids were used at 100 ng for different promoter constructs. The amount of p73α used was 2 ng for Ipaf-CAT promoter and 50 ng for MDM2-Luc promoter. The amount of p73β used was 10 ng for PG13-Luc promoter. Relative reporter activities were calculated after normalizing with β-galactosidase activities. Data presented here are mean ± S.D. of at least three independent experiments, *p < 0.01.(E) Effect of c-Src on p73 induced Ipaf promoter transctivation. HeLa cells transfected with Ipaf promoter construct along with p73α (2 ng) and c-Src or Hck (100 ng) were subjected to CAT activity measurements. Data presented here are mean ± S.D. of at least three independent experiments, *p < 0.05, **p < 0.01. (F, G) Total RNA isolated from Hela cells transfected with HA-p73α (200 ng) or Hck (1300 ng) and mSH3 Hck (1300 ng) were subjected to RT-PCR analysis for Ipaf, MDM2 and PUMA gene expression. p73α and Hck mRNA levels were determined for transfection efficiency control whereas GAPDH mRNA levels were used as an internal control.

Back to article page