Journal: Cell Death & Disease

Article Title: Lomitapide, a cholesterol-lowering drug, is an anticancer agent that induces autophagic cell death via inhibiting mTOR

doi: 10.1038/s41419-022-05039-6

Figure Lengend Snippet: a Significantly enriched pathways in lomitapide-treated HCT116 cells compared with vehicle-treated cells identified through KEEG analysis. b Volcano plot showing significant gene expression changes in response to lomitapide treatment in HCT116 cells. c HT29 cells were transfected with GFP-LC3 plasmid for 24 h, and treated with 5 μM lomitapide for another 24 h. GFP-LC3 puncta was visualized by a confocal microscope. Scale bar: 20 μm. d Cell viability was measured in HT29 cells treated with 5 μM lomitapide in the absence or presence of 100 nM bafilomycin for 24 h. e HT29 cells were treated with 5 μM lomitapide in the absence or presence of 1 mM 3-MA for 24 h. LC3 levels were measured by immunoblotting to assess autophagy induction. f si-control and siATG7–transfected HT29 cells were treated with 5 μM lomitapide for 24 h. LC3 levels were measured by immunoblotting to assess autophagy induction.

Article Snippet: About 2 × 10 5 HT29 cells were inoculated in a six-well plate and incubated at 37 °C overnight, and then siRNAs were added for 48 h. For knock-down experiments, siATG7 (SignalSilence #6604; Cell Signaling Technology), siBeclin-1 (SignalSilence #6222; Cell Signaling Technology), siAMPK (siAMPK #1: 5′-CGACUAAGCCCAAAUCUUU-3′, siAMPK #2: 5′-ACCAUGAUUGAUGAUGAAGCCUUAA-3′) were used and then cells were treated with 0 or 5 μM concentration of lomitapide at 24 h prior to the endpoint.

Techniques: Expressing, Transfection, Plasmid Preparation, Microscopy, Western Blot