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. 2018 Apr 9;128(5):1937–1955. doi: 10.1172/JCI95089

Figure 1. ZMYND8 is a direct HIF-1 and HIF-2 target gene.

Figure 1

(A) RT-qPCR analysis of ZMYND8 mRNA levels in breast cancer cells exposed to 20% or 1% O2 for 24 hours (mean ± SEM, n = 3). *P < 0.05, **P < 0.01, ****P < 0.0001 versus 20% O2, by 2-tailed Student’s t test. MDA-231, MDA-MB-231; MDA-468, MDA-MB-468. (B) Immunoblot assays of indicated proteins in breast cancer cells exposed to 20% or 1% O2 for 24 hours (n = 3). (C) RT-qPCR analysis of ZMYND8 mRNA levels in parental, HIF-1α–KO, HIF-2α–KO, and HIF-1/2α–DKO MDA-MB-231 cells exposed to 20% or 1% O2 for 24 hours (mean ± SEM, n = 3). *P < 0.05, **P < 0.01, ****P < 0.0001, by 2-way ANOVA with Tukey’s t test. DKO, double KO. (D) Immunoblot assays of indicated proteins in parental, HIF-1α–KO, HIF-2α–KO, or HIF-1/2α–DKO MDA-MB-231 cells exposed to 20% or 1% O2 for 24 hours (n = 3). (E) Nucleotide sequence of the HRE (in red) at the promoter of the ZMYND8 gene. (F) ChIP-qPCR assays in T47D cells exposed to 20% or 1% O2 for 24 hours (mean ± SEM, n = 3). *P < 0.05, ****P < 0.0001 versus 20% O2, by 2-way ANOVA with Sidak’s t test. (G–I) Luciferase reporter assays in HEK293T (G and H) and HeLa (I) cells transfected with the indicated plasmids and exposed to 20% or 1% O2 for 24 hours. The FLuc/RLuc activity was determined (mean ± SEM, n = 3). **P < 0.01, ***P < 0.001, ****P < 0.0001, by 2-way ANOVA with Tukey’s t test. Mut, mutant.