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. 2021 May 25;24(6):102649. doi: 10.1016/j.isci.2021.102649

Figure 1.

Figure 1

KEAP1 deficiency promotes glucose dependency in lung cancer cells mainly through SLC7A11

(A) Protein levels of KEAP1, NRF2, and SLC7A11 in different NSCLC cell lines were determined by Western blotting. Vinculin is used as a loading control.

(B) Cell death upon glucose (Glc) starvation for 0–24 hr in different NSCLC cell lines was analyzed by PI staining.

(C) Western blotting analysis of KEAP1, NRF2, and SLC7A11 protein levels in the control (sgC) and KEAP1 knockout (sg1/2/3) H1299 cells.

(D) Cell death in KEAP1 knockout H1299 cells upon glucose withdrawal was analyzed by PI staining.

(E) Western blotting analysis of KEAP1, NRF2, and SLC7A11 protein levels in KEAP1-overexpressing H460 cells. EV, empty vector, OE, KEAP1 overexpressing.

(F) Cell death in KEAP1-overexpressing H460 cells upon glucose withdrawal was analyzed by PI staining.

(G) Western blotting analysis of KEAP1, NRF2, and SLC7A11 protein levels in NRF2-KEAP1 double knockout H1299 cells.

(H) Cell death in NRF2-KEAP1 double knockout H1299 cells upon glucose withdrawal was analyzed by PI staining.

(I) Correlation analysis of SLC7A11 expression in KEAP1 WT vs mutant LUAD tumors from TCGA.

(J) Correlation analysis of SLC7A11 expression with KEAP1 mutation status of NSCLC in CCLE database.

(K) Western blotting analysis of KEAP1, NRF2, and SLC7A11 protein levels in SLC7A11-KEAP1 double knockout H1299 cells.

(L) Cell death in SLC7A11-KEAP1 double knockout H1299 cells upon glucose withdrawal was analyzed by PI staining.

(M) Cell death in SLC7A11-KEAP1 double knockout H1299 cells upon glucose withdrawal supplemented with erastin was analyzed by PI staining.

(N) Western blotting analysis of SLC7A11 protein levels in SLC7A11 knockdown (sh1/2) H460 cells.

(O) Cell death upon glucose withdrawal was analyzed by PI staining in SLC7A11 knockdown H460 cells.

(P) SLC7A11 protein levels in SLC7A11-overexpressing H1299 cells. EV, empty vector, OE, SLC7A11 overexpressing.

(Q) Cell death in SLC7A11-overexpressing H1299 cells upon glucose withdrawal was analyzed by PI staining.

Data are represented as mean ± SD; n = 3. ∗, p < 0.05, ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001.