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. 2020 Nov 2;9:e60151. doi: 10.7554/eLife.60151

Figure 6. Mutant KRAS and Myc promotes p-4E-dependent glutamine addiction.

Indicated WT and mutant KRAS isogenic CRC lines were subject to glutamine deprivation and analyzed. (A) The basal levels of indicated proteins were analyzed by western blotting. (B) Attached cells at 48 hr were stained with crystal violet. (C) Cell death at 48 hr was quantitated by flow cytometry. Representative results were shown. (D) The indicated proteins at 24 hr in two pairs of cell lines were analyzed by western blotting. *, non-specific band. (E) Heat map of indicated transcripts at 24 hr were detected by RT-PCR and normalized to the control (WT, Gln+, 1). (F) Numbers of differential genes upon Gln deprivation in the isogenic HCT 116 pair, 2-fold or more, *p<0.05. (G) Venn diagram of induced genes from (F). (H–J) Reactome pathway analysis of (H) shared, WT (I), and 4EKI-specific upregulated genes(J). Top five enriched pathways are shown.

Figure 6.

Figure 6—figure supplement 1. Mutant KRAS promotes glutamine addiction.

Figure 6—figure supplement 1.

Two pairs of wt and mutant KRAS isogenic CRC cells were subjected to glutamine deprivation. (A and B) Cells at 48 hr were analyzed by flow cytometry with quantification of PI+ and PI- AnnexinV+ populations. (C) The indicated proteins at 24 hr were analyzed by western blotting. (D) Scatter plot of microarray data from isogenic HCT 116 cells 24 hr after Gln deprivation.