Fig. 3.

PHF8 is essential for maintaining H3K4me3 levels on hypoxia-inducible genes. A. Schematic illustration of PCR amplicons for probing the proximal promoter region (Amp 1) or non-specific control loci (Amp 2) of the indicated genes. B. LNCaP cells stably expressing doxycycline-inducible control (Ctrl) or PHF8 shRNAs were cultured under normoxia (Nor) or hypoxia (Hyp; 1% O₂) for 12 h. The specific ChIP antibodies are indicated by row. The ChIP-PCR data are interpreted as percentage of 1% input DNA, with the values between amplicons compared as follows: 1) in control RNAi cells, Amp 1 versus Amp 2 under normoxia and under hypoxia; 2) in PHF8 RNAi cells, Amp 1 versus Amp 2 under normoxia and under hypoxia; 3) Amp1 between control and PHF8 RNAi cells under normoxia and under hypoxia, the same amplicon 2.

The S.D. was obtained from at least three independent experiments. For calculation of significance, two-way ANOVA was used in the case of multiple comparisons, and the Student’s t-test was used in the case of paired comparison(s). *: p < 0.05; **: p < 0.01.