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. 2017 Sep 14;14:164–177. doi: 10.1016/j.redox.2017.08.022

Fig. 5.

Fig. 5

OGC downregulation promotes hypoxia-mediated CLOOH, which reverses cholesterol-mediated inhibition of membrane permeabilization. A, Representative kinetics of BAX+tBID induced FD70 release from vesicle LUV composed of PC/CL (-CHOL-CLOOH), PC/CL/CHOL (+CHOL-CLOOH), or PC/ CLOOH/CHOL (+CHOL+CLOOH). BAX and tBID concentrations were 40 nM, and dose-dependence effects of CLOOH on LUV permeabilization induced by different pore-forming proteins in LUV composed of PC/CL/CHOL in which CL was substituted by indicated molar amounts of CLOOH. Concentrations of BAX, BAKΔC, tBID, and tetanolysin were 40 nM, 40 nM, and 3 nM, respectively. Data represent mean values and standard errors of 3–6 independent measurements. B-C, CHOL and CLOOH do not significantly affect BAX recruitment to liposomes (B) nor BAX oligomerization (C). BAX, monomeric BAX; Multi-BAX, oligomeric BAX. D, CLOOH reverses the inhibitory effect exerted by CHOL on the capacity of BAX to penetrate into lipid monolayers spread at 30 mN/m followed by addition of octylglucoside-oligomerized BAX (500 nM) into the subphase, and monolayer surface pressure increase was determined when the signal reached a plateau. Data represent mean values and S.E. of 3–4 independent experiments. E, CLOOH reverses the increase in the micropolarity of the lipid bilayer hydrocarbon core exerted by CHOL determined by polarization fluorescence emission spectra of indicated multilamellar vesicles containing 5 μM DPH incorporated. Excitation wavelength was 370 nm. All spectra were corrected by substracting the spectra of buffer, or spectra of phospholipid suspensions without fluorescent probe. Representative fluorescence emission spectra of 0.2 mol% F-DHPE incorporated in multilamellar vesicles of indicated lipid composition. Excitation wavelength was 480 nm. All spectra were corrected by substracting the spectra of buffer, or spectra of phospholipid suspensions without fluorescent probe. F, Representative thermograms obtained by differential scanning calorimetry for multilamellar vesicles of indicated lipid compositions. MirCL, 50 mol miristoyl cardiolipin plus 50 mol PC, a lipid mixture used as a positive control for lipid domain formation.