Fig. 4

Formo reduces Mcl-1 protein level and induces apoptosis in NSCLC cells. a, z-VAD-fmk compromised Formo decreased cell viability. HCC827, H1975, and A549 cells were pre-treated with z-VAD-fmk, necrostatin-1, or GSK’872 for 2 h, followed by Formo (10 μM) treatment for 24 h, cell viability was analyzed by MTS assay. b-d, Formo promotes apoptosis in NSCLC cells. NSCLC cells were treated with Formo for 24 h, WCE was subjected to IB analysis (b), and caspase 3 activity analysis (c). Flow cytometry analysis was performed to examine the population of apoptotic cells (d). e, HCC827 cells were treated with Formo for 24 h, subcellular fractions were isolated and subjected to IB analysis. Cyto, cytoplasmic fraction; Mito, Mitochondrial fraction. f, NSCLC cells were treated with Formo for 24 h, WCE was subjected to IB analysis. g, HCC827 cells were transfected with Mcl-1 and treated with Formo for 24 h. Subcellular fractions were isolated and subjected to IB analysis. H-K, HCC827 cells were transfected with Mcl-1 and treated with Formo for 24 h, the cell viability was analyzed by MTS assay (h), live cell population was determined by trypan blue exclusion assay (i). WCE was prepared and subjected to caspase 3 activity analysis (j). The apoptotic cell was determined by flow cytometry (k). C-PARP, cleaved-PARP; C-caspase 3, cleaved- caspase 3