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. 2015 Dec 31;113(3):644–649. doi: 10.1073/pnas.1517112113

Fig. S4.

Fig. S4.

FAM129B Y593 phosphorylation promotes the Warburg effect and tumor cell proliferation and invasion. (A) U87/EGFRvIII cells depleted or not depleted of FAM129B and with reconstituted expression of WT rFAM129B or rFAM129B Y593F mutant (Left) were cultured in no-serum DMEM for 18 h. The media were collected for analysis of glucose consumption (Middle) or lactate production (Right). Data represent the means ± SD of three independent experiments. Immunoblotting analyses were performed with the indicated antibodies. (B) U87/EGFRvIII cells depleted or not depleted of FAM129B and with reconstituted expression of WT rFAM129B or rFAM129B Y593F mutant were cultured for 5 d and harvested for cell counting. Data represent the mean ± SD of three independent experiments. (C) U87/EGFRvIII cells depleted or not depleted of FAM129B and with reconstituted expression of WT rFAM129B or rFAM129B Y593F mutant were plated on the surface of Matrigel coating the upper chamber of Transwell chambers. After 16 h, cells that migrated to the opposite side of the insert were stained with crystal violet. The membranes with invaded cells were dissolved in 4% (wt/vol) deoxycholic acid and read colorimetrically at 590 nm for quantification of invasion. Data represent the mean ± SD of three independent experiments.