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Fig. 4 | Molecular Cancer

Fig. 4

From: Regulation of stem-like cancer cells by glutamine through β-catenin pathway mediated by redox signaling

Fig. 4

Glutamine depletion leads to a reduction in cellular GSH and ROS accumulation. a A549 cells cultured in complete were switched to medium without glutamine for the indicated time. The cells were collected for analysis of intracellular GSH content as described in Materials and Methods. b Determination of cellular ROS in A549 cells. Cells were cultured as in (a), and ROS was detected by flow cytometry using DCF-DA. c Quantitation of cellular ROS under conditions described in (b). Each bar represents mean ± SD, n = 3. d Determination of superoxide in A549 cells cultured with or without glutamine for the indicated time, cellular superoxide was detected by flow cytometry using Het staining. e A549 cells were cultured in glutamine-free medium or in complete medium and treated with L-Asparaginase for the indicated time, NADP+/NADPH ratio was then determined as described in Materials and Methods. f Comparison of cellular GSH contents in sorted SP and non-SP cells. g Comparison of expression of genes involved in synthesis of glutathione in sorted SP and non-SP cells. GCLC, γ-glutamylcysteine ligase catalytic subunit; GCLM, γ-glutamylcysteine ligase modulatory subunit; GSS, glutathione synthetase. Each bar represents the mean of the relative ratio ± SD, n = 3; *, p < 0.05; **, p < 0.01

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