Fig. 6 | Signal Transduction and Targeted Therapy

Fig. 6

From: RBM4 dictates ESCC cell fate switch from cellular senescence to glutamine-addiction survival through inhibiting LKB1-AMPK-axis

Fig. 6The alternative text for this image may have been generated using AI.

Glutamine metabolism was elevated by RBM4-LKB1 axis to promote ESCC cells survival. a Heatmap analysis of metabolism-related genes upon RBM4 depletion or overexpression in KYSE150 cells. b, c Glutamine consumption and glutamate levels were examined in KYSE30, KYSE150 and KYSE450 cells with stable overexpression of RBM4 (b) or depletion of RBM4 (c). Three experiments were performed and mean ± SD was plotted. P-values were determined by Student’s t-test (b) or One-way ANOVA with Dunnett multiple comparisons (c). d The glutamate production was examined in KYSE150 and KYSE450 cells with stable overexpression of RBM4 or depletion of RBM4 while glutamine deprivation. Three experiments were performed and mean ± SD was plotted. P-values were determined by Student’s t-test or One-way ANOVA with Dunnett multiple comparisons. e, f KYSE150 and KYSE450 cells with stable overexpression of RBM4 (e) or depletion of RBM4 (f) were subjected to measure the reduced GSH/ oxidized GSSG ratio calculated by GSH/GSSG = [Total GSH-(2 × GSSG)]/GSSG. Three experiments were performed and mean ± SD was plotted (P-values were determined by One-way ANOVA with Dunnett multiple comparisons or Student’s t-test). g The growth curve of RBM4-overexpressed KYSE30, and KYSE150 cells with or without CB-839 treatment was measured by CCK8 assay. P-values were determined by two-way repeated measures ANOVA. h Glutamine consumption and glutamate levels of RBM4-depleted KYSE150 cells in the presence or absence of LKB1 were examined. Three experiments were performed with mean ± SD plotted (P-values were determined by One-way ANOVA with Dunnett multiple comparisons). i Glutamine consumption and glutamate levels of RBM4-depleted KYSE150 cells in the presence or absence of AMPK inhibitor compound C (5 μM) were examined. Three experiments were performed with mean ± SD plotted (P-values were determined by One-way ANOVA with Dunnett multiple comparisons). j β-gal staining of KYSE150 cells with stable knockdown of RBM4 in the presence or absence of glutamic acid (0.5 mM), methyl pyruvate (10 mM) and NAC (3 mM). Three experiments were carried out with mean ± SD of β-gal positive cells plotted (P-values were determined by One-way ANOVA with Dunnett multiple comparisons). Scale bar = 25 μm. k The growth curve of RBM4-depleted KYSE150 cells with or without addition of glutamic acid, methyl pyruvate, and NAC was measured by CCK8 assay. P-values were determined by two-way repeated measures ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

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