Fig. 5: PAMT-001-mediated autophagy contributes to apoptotic cell death.
From: A novel PAAoptosis-inducing ERRα-targeting compound for combating hematopoietic and solid cancers
A Western blotting for PAMT-001-induced autophagic flux in HL-60 (left) and HCT-116 (right) cells. B Representative TEM images of PAMT-001-treated KG1α (left) and HCT-116 (right) cells. Multiple double-membrane autophagosomes (red arrow) or mitophagosomes (yellow arrow) accumulated in the cytoplasm. C, D Cytotoxicity analysis for detecting the rescue of PAMT-001-induced cell death by autophagy inhibitor (bafilomycin A1 and chloroquine) using trypan blue exclusion assay for HL-60 cells, CCK-8 assay for THP-1 cells, and MTT assay for primary AML #125, HCT-116, and HT-29 cells. Cells were cotreated with PAMT-001 and bafilomycin or chloroquine for 24 h (HL-60 and THP-1), 48 h (primary AML cell #125), or 30 h (HCT-116 and HT-29). P-values were calculated using one-way ANOVA for multiple comparisons (mean ± SD). E Optical microscopy images showing PAMT-001-induced HL-60 cell death and its rescue by bafilomycin A1. F Western blotting for autophagy-dependent cell death in KG1α (left) and HCT-116 cells(right). Expression of apoptotic markers (cleaved caspase-3 and caspase-9) and CHOP was attenuated by bafilomycin A1 treatment. G MTT assay for detecting ATG7-dependent cell death. HT-29 with ATG7 knockout (KO) cells showed more resistance to PAMT-001-induced cell death than those with ATG7 wild type. P-values were calculated using a two-sided t-test (mean ± SD, n = 6). ***P < 0.001, ****P < 0.0001.
