Fig. 3: BMAL1 and HIF2α form an active heterodimer.
From: BMAL1 and ARNT enable circadian HIF2α responses in clear cell renal cell carcinoma
A Heparin chromatography elution of BMAL1 and HIF2α co-expressed in insect cells. SDS-PAGE analysis shows a co-eluted stoichiometric complex of BMAL1-HIF2α from one of two independent experiments with similar results. B Mass photometry of purified BMAL1-HIF2α complex. A minor peak centered at 91 kDa corresponds to the molecular weight of HIF2α, suggesting that it is in slight excess. The major peak, centered at 157 kDa, is consistent with the calculated molecular weight for the BMAL1-HIF2α heterodimer. C Detection of endogenous HIF2α and CLOCK and of ectopically expressed FLAG-tagged class I bHLH-PAS proteins by immunoblot of lysates (input) or complexes purified with an anti-FLAG antibody from 786O cells expressing the indicated plasmids. The data represent one of three independent experiments with similar results. D Relative luminescence units detected in HEK293T cells expressing luciferase under the control of a hypoxia-responsive element with the indicated additional plasmids. ****P < 0.0001 by one-way ANOVA with Tukey’s correction for multiple comparisons. Data in (D) depict the mean ± s.d. for n = 5 biological replicates from one of three independent experiments with similar results. Source data are provided as a Source Data file.
