Extended Data Fig. 1: GS is required for cancer cell proliferation in a glutamine-sufficient condition.
From: Glutamine synthetase licenses APC/C-mediated mitotic progression to drive cell growth
a, Quantification of cell proliferation of H460 cells transduced with shRNAs targeting 35 amino acid metabolic enzymes. The n of shRNAs used for each gene was shown as indicated and in Supplementary Table 1. b, Knockdown (KD) efficacy of shRNAs for the indicated enzymes. (3–5 individual shRNA per gene; shown was the mean of 3 independent biological replicates). c, Expression of GS in the human lung and liver cancer cell lines transduced with control or GS-targeted shRNAs. d, Expression of GS in H460 cells treated with or without MSO in the presence of 2 mM glutamine. e, Expression of mouse GS (mGS) in the mouse lung and liver cancer cell lines transduced with control or mGS-targeted shRNAs. f, Cell proliferation of KP cells transduced with control shRNA or shmGS-1 (left) or treated with solvent control or 1 mM MSO (right) in the presence of 2 mM glutamine. g, Colony formation by the indicated cells in the presence of 2 mM glutamine. h, Cell colonies formed by indicated cells treated with solvent control or 1 mM MSO in the presence of 2 mM glutamine. i, Expression of GS in A549 cells transfected with control siRNA or pooled siGS oligos. j, Cell growth of A549 cells transfected with control siRNA or pooled siGS oligos for 5-day culture with 2 mM glutamine. (n=3 independent biological replicates for f-g, j). k, Expression of GS in two patient-derived organoids transduced with control shRNA or GS-targeted shRNAs. l, Luc2-transduced Huh7 cells were intrahepatically implanted into nude mice. AAV expressing control (shScram) or GS-targeted (shGS) hairpins was delivered by tail vein injection and the expression of GS was measured by western blots. m, Luc2-transduced Huh7 cells were intrahepatically implanted into nude mice, and animals were treated with PBS or MSO (20 mg/kg) daily for two weeks. Tumor growth was monitored by measuring luciferase activity. (n=5 for each group of tumors). Data were presented as mean±s.e.m. Unpaired, two-tailed t-tests were applied for f, g, h, j, and m. *: p<0.05, **: p<0.01.
