Fig. 4: PAMT-001 promotes cell death by stimulating ER-stress-related genes, especially CHOP.
From: A novel PAAoptosis-inducing ERRα-targeting compound for combating hematopoietic and solid cancers
A, B GSEA against GOBP: Cellular response to unfolded protein (B, left) and GOBP: Response to endoplasmic reticulum stress Genes (B, right) were ranked based on fold changes between vehicle and PAMT-001 treated KG1α cells. Quantification of gene expression of ER stress-related gene (CHOP, BiP, EDEM, and ATF4) was measured by qRT-PCR at the indicated concentration and time in KG1α (C), primary AML cell (D), and A549 (E). Of these, PAMT-001 primarily increased CHOP expression. P-values were calculated by a two-sided t-test (mean ± SD, n = 3, respectively). The western blot analysis of PAMT-001-induced CHOPA expression in HCT-116 (F) and HT-29 (G). H Quantification of CHOP expression induced by PAMT-001 (2.5 μM, 24 h) in HL-60 with transfection of siRNA targeting CHOP (siCHOP) and non-targeting (siNS). P-values were calculated by one-way ANOVA for multiple comparisons (mean ± SD, n = 3, respectively). I The LD50 value of PAMT-001 according to CHOP expression in HL-60 cells. The LD50 values were calculated through [inhibitor] vs. normalized response—Variable slope in Prism 8. P-values were calculated by the extra sum of squares F-test. J The cytotoxicity analysis for detecting rescues PAMT-001-induced cell death by ER stress inhibitor (4-PBA; 4-Phenylbutyric acid) using CCK-8 assay in HL-60. P-values were calculated by one-way ANOVA for multiple comparisons (mean ± SD). ****P < 0.0001.
