Fig. 6: EI-52 induces immunogenic cancer cell death in vitro.
From: Targeting ERK-MYD88 interaction leads to ERK dysregulation and immunogenic cancer cell death
A RNAseq GSEA plot and (B) top upregulated genes in HCT116 cells treated for 18 h with EI-52 (6 μM) or DMSO. C Chemokine mRNA from HCT116 cells treated for 16 h with DMSO or EI-52 8 μM. Mean ± SEM from three independent experiments, two-tailed one-sample t-test. CXCL8 **p = 0.0029; CXCL1 *p = 0.0312; CXCL2 *p = 0.0439 (95% confidence interval). D CXCL8, CXCL1, and CXCL2 in supernatants of HCT116 cells treated with DMSO, MEK inhibitor U0126 10 μM, or EI-52 8 μM for 24 h. Mean ± SEM from four (CXCL8) and three (CXCL1, CXCL2) independent experiments, unpaired two-tailed Student’s t-test; CXCL8 **p = 0.0073; CXCL1 *p = 0.0479; CXCL2 *p = 0.0325 (95% confidence interval) (E) NF-κB, CXCL1, and CXCL8 luciferase activity 16 h post-treatment with EI-52 8 μM or DMSO. Mean of treatment /DMSO ratio ± SEM from four (CXCL8) and three (CXCL1, CXCL2) independent experiments, unpaired two-tailed Student’s t-test; NF-κB *p = 0.0395; CXCL1 *p = 0.0165; CXCL8 *p = 0.0108 (95% confidence interval). F IL8 secretion in supernatant of HCT116 cells transfected for 24 h with control or p65 siRNA, then treated 16 h with DMSO or EI-52 8 μM. Mean ± SEM from three independent experiments, unpaired two-tailed Student’s t-test; ***p = 0.0002 (95% confidence interval) (G) Co-immunoprecipitation of ERK1-HA with p65-Flag after treatment of transfected HEK293T cells with EI-52 8 μM. Representative data of three independent experiments. H Extracellular ATP and HMGB1 in supernatants of HCT116 cells treated for 24 h with DMSO, U0126 10 μM, or EI-52 8 μM. Results are expressed as mean of treatment/DMSO ratio ± SEM from four (e-ATP) or three (HMGB1) independent experiments, two-tailed one sample t-test; e-ATP: U0126 *p = 0.0171; EI-52 **p = 0.0034 (95% confidence interval). HMGB1: EI-52 *p = 0.0157 (95% confidence interval). I Chemotaxis of migrating THP1 cells was evaluated in Boyden chamber containing supernatant of HCT116 cells treated 24 h with control, U0126 10 μM, or EI-52 8 μM. Mean ± SEM from three independent experiments, Dunnett’s one-way ANOVA; **EI-52 p = 0.0013 (95% confidence interval). Source data are provided as a Source Data file.
