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. 2015 Feb 20;11(2):253–270. doi: 10.1080/15548627.2015.1009778

Figure 3.

Figure 3.

SIRT5 controls glutamate accumulation. (A) MDA-MB-231 and C2C12 WT cells, either treated with MC3482 or left untreated, as well as SIRT5+ and SIRT5- cells were processed to obtain whole cellular extracts. Glutamate concentration was measured using a glutamate assay kit according to manufacturer's protocol. Alternatively, glutamate was also measured in MDA-MB-231cells treated with the GLS inhibitor BPTES for 17 h. Data are representative of at least 3 separate experiments. *Significantly different from WT cells. Significance was set at P < 0.05. (B) MDA-MB-231 and C2C12 WT cells in the presence or absence of MC3482, as well as SIRT5+ and SIRT5- clones were kept in culture for the times indicated. In addition cells were also treated with the GLS inhibitor BPTES or with dimethyl-α-ketoglutarate to replenish anaplerotic flux. Ammonia levels were measured in the culture medium after 2 and 3 d as reported under Materials and Methods. Data are representative of at least 3 separate experiments. *Significantly different from nontreated (NT) cells. Significance was set at P < 0.05.