Reducing hLgl levels increases cell survival under hypoxia. (A) Protein levels of hLgl-1 are severely reduced in hLgl-KD1 and hLgl-KD2 stable lines expressing hLgl-1 shRNA, but not in the control cells that express scrambled shRNA. αTubulin (αTub) is the loading control. (B) Cell death in control, hLgl-KD1, and hLgl-KD2 cells incubated under normoxia or hypoxia (1% O2/5%CO2), respectively. Live cells are labeled by calcein AM (green), and dead cells are labeled by ethidium homodimer-1 (red). Cell death decreases after day 1 likely because old dead cells were lost by medium washes, which are required before dye staining. (C) Quantification of the cell viability after normoxia or hypoxia exposures in stable HEK293 cell lines expressing hLgl-1 shRNA (hLgl-KD1 and hLgl-KD2) or scrambled shRNA (Control) as shown in B. *, P < 0.05; **, P < 0.01; ***, P < 0.001. ns, not significant. Bars, 5 µm. Error bars represent means ± SEM.
