Figure 3.

p53–HIF-1α complex is inhibited by TGase 2 inhibition in hypoxia. (A) ACHN cells were treated with STN for 2 h and incubated for 4 h in hypoxia (1% O₂). The proteins were immunoprecipitated from cell extracts using an anti-p300 antibody and subjected to immunoblotting. (B) The image J analysis of Western blotting of Figure 3A. (C) Cells were transfected with 3xFLAG p53 (p53 overexpression vector; 0, 2, 4 μg) for 8 h and incubated for 4 h in hypoxia. The total RNAs were isolated from cells and RT-PCRs against mouse double minute 2 homolog (MDM2), TGM2, and GAPDH were performed as described in the method. GAPDH was used as an internal control for the RT-PCR system. Error bar represents SD. GraphPad Prism software used to perform one-way ANOVA, ns = not significant. Data are representative of three independent experiments.