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. Author manuscript; available in PMC: 2017 Jan 24.
Published in final edited form as: Mol Cancer Res. 2012 May 1;10(7):937–944. doi: 10.1158/1541-7786.MCR-11-0493-T

Figure 4.

Figure 4

PA200 is required to maintain intracellular free glutamine pools. A, after transfection, HeLa cells were immediately plated (10,000 cells per well of 96-well plate) in DMEM (which lacks glutamate) containing or lacking 10% bovine calf serum, 2 mmol/L L-glutamine, and 4.5 g/L glucose. Five days later, cell growth/metabolic activity was measured using the Promega cell proliferation kit (MTS). B, HeLa cells were transfected with PA200-specific siRNA (white bars), control siRNA (black bars), or PA200-specific siRNA together with PA200 cDNA that is insensitive to the siRNA (gray bars). After media replacement on day 2 after transfection, cells were allowed to grow for 3 more days. Deproteinized cell pellets (B) were analyzed for the indicated amino acids using the PicoTag method. Supernatants (C) were analyzed for glutamate remaining in the media using a BioProfile FLEX (Nova Biomedical). A representative figure of at least 3 independent experiments is shown for each panel.