Characterization of the effects of Haps59 expression in Flp-In 293 cells and <i>E. coli</i>.

<p>(A) Western blot with anti-HIF-1α antibodies showing that the addition of CoCl₂ causes the accumulation of HIF-1α in Flp-In 293 cells. See <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114032#pone.0114032.s005" target="_blank">Figure S5</a> for full blot. (B) Toxicity of 5FC to stable isogenic pools of Flp-In 293 cells containing EV, yCD, and Haps59. Cells were grown in the absence (i.e. −HIF-1α) or presence (i.e. + HIF-1α) of 50 µM CoCl₂, and increasing concentrations of 5FC. Percent survival was calculated by measuring the total DNA of surviving cells in each well and normalizing between wells containing no 5FC (i.e. 100% survival) and wells containing no cells (i.e. 0% survival). Error bars represent the standard deviation across three replicates. (C) Dot toxicity assay indicates that CITED2 but not E1A activates Hap59 in <i>E. coli</i>. Equal density dilutions of log phase cultures of strains expressing Haps59 and either the p300 binding fragments of HIF-1α, CITED2, or E1A were plated on increasing concentrations of 5FC.