HIF1α and HIF2α ChIP in VSMCs

Hif1a^fl/fl or Hif1a^ΔSMC VSMCs treated with Ang II (1 μM) for 24 h were crosslinked in 1% formaldehyde in 1× PBS for 10 min. ChIP assays were performed for HIF1α (2 μg/IP, NB100-105; Novus Biologicals) or HIF2α (2 μg/IP, NB100-122; Novus Biologicals) using Simple ChIP Plus Kit (Cell Signaling Technology, Danvers, MA, USA) as previously described^{14 (link)}. The primers for ChIP assays are listed in Supplementary Table 1.

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Qi D., Wei M., Jiao S., Song Y., Wang X., Xie G., Taranto J., Liu Y., Duan Y., Yu B., Li H., Shah Y.M., Xu Q., Du J., Gonzalez F.J, & Qu A. (2019). Hypoxia inducible factor 1α in vascular smooth muscle cells promotes angiotensin II-induced vascular remodeling via activation of CCL7-mediated macrophage recruitment. Cell Death & Disease, 10(8), 544.

Publication 2019

NB100-105 NB100-122 Simple ChIP Plus Kit

Biologicals Chip Chip assays Formaldehyde Hif1α Novus Primers

Corresponding Organization :

Other organizations : Ministry of Education of the People's Republic of China, Capital Medical University, University of Michigan–Ann Arbor, Dalian Medical University, First Affiliated Hospital of Dalian Medical University, King's College Hospital, Beijing Anzhen Hospital, National Cancer Institute, National Institutes of Health, Center for Cancer Research

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Variable analysis

independent variables

Hif1a floxed (Hif1a^fl/fl) or Hif1a knockout (Hif1a^ΔSMC) VSMCs
Angiotensin II (Ang II) treatment (1 μM) for 24 h

dependent variables

HIF1α and HIF2α binding to DNA, as measured by ChIP assays

control variables

Formaldehyde crosslinking concentration (1%)
Crosslinking duration (10 min)
Buffer used for crosslinking (1× PBS)

controls

Positive control: Antibodies used for ChIP assays (HIF1α and HIF2α)
Negative control: Not explicitly mentioned

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