Figure 2
Knockdown of endogenous ME2 induces A549 cell death and differentiation and inhibits ATP and NADPH production, while enhancing A549 cells oxygen consumption and ROS production.
(a), Depletion of ME2 increases A549 basal cell death. (b), E-cadherin, ZO-1, ME2 and β-tubulin in A549 cells with or without ME2 knockdown were determined by Western blotting. The full-length blots are presented in Supplemental Figure S10. Data are representative of two independent experiments. (c), Depletion of ME2 inhibits ATP production in A549 cells. Data are expressed as mean ± SD, n = 3. (d), Knockdown of ME2 in A549 cells increases O2 consumption. (e), Knockdown of ME2 in A549 cells increases [H+] excretion. (f), Increased 13C3-lactate excretion induced by ME2 knockdown in A549 cells, as determined by 1H NMR; data are expressed as mean ± SEM, n = 2. (g), Increased ROS in A549 ME2 knockdown cells detected by flow cytometry using CM-H2DCFDA as probe. Each histogram is representative of three experiments. (h), 5 mM exogenous NAC inhibits ME2 depletion-meditated ROS production. Each histogram is representative of two or three experiments. (i), 5 mM NAC partially rescues ME2 knockdown induced cell death. Control groups without NAC treatment were normalized to 100 and NAC treated groups were compared to its control. Data are expressed as mean ± SD, n = 3. (j), Depletion of ME2 increases NADP/NADPH ratio. Data are expressed as mean ± SD, n = 3.
