Extended Data Fig. 2: HIF1α stabilization is independent of hypoxia during CHP stimulation.
From: Mechanosensation of cyclical force by PIEZO1 is essential for innate immunity
a, Representative confocal microscopy of BMDMs treated for 6 h in either normoxic conditions or in a hypoxic chamber (2% O2). Cells were treated with pimonidazole (20 μM) for 1 h before completion of treatment. Upon completion, cells were fixed, permeabilized and stained with monoclonal antibody against pimonidazole. Data are representative of two independent experiments. b, Confocal microscopy of BMDMs treated for 6 h in either static, CHP or hypoxic chamber (0.5% O2). Cells were treated with 20 μM pimonidazole 1 h before completion of treatment. Upon completion, cells were fixed, permeabilized and stained with monoclonal antibody against pimonidazole. Data are representative of two independent experiments. c, RT–qPCR analysis of BMDMs treated with cyclical hydrostatic pressure, 2% O2 or cyclical hydrostatic pressure with 2% O2 for 6 h. Data are presented as three biological replicates. d, RT–qPCR analysis of Hif1a in BMDMs following CHP. Data are presented as four biological replicates. Data are from two independent experiments. e, Representative immunoblot analysis of HIF2α and β-tubulin from BMDMs treated with static pressure, CHP or DMOG (200 μM) for 6 h. Data in c, d are mean ± s.e.m.; significance is determined by unpaired two-tailed t-test.
