PM targeting of mLgl::GFP in MDCK cells under hypoxia and reoxygenation. MDCK cells cultured for 3 d in MEMα medium on 12-mm diameter filters (pore size, 0.4 µm) were mounted and imaged in custom micro-imaging chambers under a steady temperature of 37°C. Flow of hypoxic gas mixture of 1% O2/5% CO2/94% N2 into the imaging chamber starts at 0 s and persists until 2,340 s. Gas flow was then switched to normal air until the end of recording. Images were analyzed by time-lapse confocal microscopy using a laser-scanning confocal microscope (LSM 510; Carl Zeiss). Frames were taken every minute for 39 min under hypoxia and every 15 s for 19 min after hypoxia.
