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. Author manuscript; available in PMC: 2021 May 30.
Published in final edited form as: Nat Metab. 2020 Nov 30;2(12):1401–1412. doi: 10.1038/s42255-020-00316-0

Figure 5. KL cells require GFPT2.

Figure 5.

a and b, Sensitivity to GFPT1 (a) and GFPT2 (b) silencing in K and KL cells (n = 6). c, Rescue effect of GlcNAc supplementation on GFPT2 silencing-induced cell death (n=6). d and e, Effects of a Dox-induced GFPT2 sgRNA (GFPT2) on anchorage independent growth of H460 (d) and H157 (e) (n=4 for H460, n = 3 for H157). NC is a Dox-inducible control sgRNA. f, Effects of Dox-induced GFPT2 KO on invasion capacity of H460 and H157 cells (n=4 for H460-sgNC−/+ Dox, n=8 for H460-sgGFPT2−/+Dox, n=12 for H157 GFPT2-Dox and H157-NC-Dox, n=8 for H157 GFPT2+Dox, n=7 for H157-NC+Dox). g, Abundance of GFPT1 and 2 in parental and GFPT2 KO H460 cells. Actin was used as a loading control. h, Effects of GFPT2 KO (two clones) on anchorage independent growth of H460. i, Effect of Dox-inducible GFPT2 KO H460 cells on cell surface L-PHA lectin binding. j, Abundance of hexosamine metabolites in Dox-inducible GFPT2 KO H460 cells (n=3). sgNC is used as a Dox-inducible sgRNA control. k, Effect of LKB1 on GFPT2 silencing-induced loss of viability (n=5). l, Effect of constitutively active AMPK (CA AMPK) on GFPT2 silencing-induced loss of viability (n=6).In b, d, e, f, h, i, j, and l, statistical significance was assessed using two-tailed Student’s t-test. In c and k, statistical significance was assessed using one-way ANOVA followed by Tukey’s post hoc test was used. In c, *, p<0.05 comparing to sieGFP without GlcNAc; #, p<0.05 comparing to sieGFP with GlcNAc treatment; $, p<0.05 comparing to siGFPT2 with GlcNAc treatment. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. Western blot was repeated three times and all other experiments were performed twice.