Journal: Cancer Science

Article Title: Peneciraistin C induces caspase‐independent autophagic cell death through mitochondrial‐derived reactive oxygen species production in lung cancer cells

doi: 10.1111/cas.12253

Figure Lengend Snippet: Peneciraistin C (Pe‐C) induces caspase‐independent non‐apoptotic cell death in three lung cancer cell lines. (a) A549, H446, H661, and BEAS‐2B cells were treated with indicated concentrations of Pe‐C for 24 and 48 h. Cell viabilities were determined by MTT assay. The IC 50 value was calculated by an IC 50 software program. (b) A549, H446, and H661 cells were treated with indicated concentrations of Pe‐C and 5‐fluorouracil (5‐FU) in the absence (–) or presence of pan‐caspase inhibitor z‐VAD‐fmk (100 μM) for 48 h. The percentage of cell death was quantified by Trypan blue staining. (c) A549 cells were treated with indicated concentrations of Pe‐C or 5‐FU for the indicated times. The activity of caspase‐3, ‐8, and ‐9 was measured by a microplate reader at 405 nm. (d) A549 cells were treated with indicated concentrations of Pe‐C or 5‐FU for 24 and 48 h. Cells were stained with propidium iodide and analyzed by flow cytometry. Data represent results of at least three independent experiments.

Article Snippet: The activity of caspase‐3, ‐8, and ‐9 was measured using commercially available caspase colorimetric assay kits (Beyotime).

Techniques: MTT Assay, Software, Staining, Activity Assay, Flow Cytometry

Journal: Cancer Science

Article Title: Peneciraistin C induces caspase‐independent autophagic cell death through mitochondrial‐derived reactive oxygen species production in lung cancer cells

doi: 10.1111/cas.12253

Figure Lengend Snippet: Blockage of autophagy converts peneciraistin C (Pe‐C)‐induced cell death to apoptotic cell death. A549 cells were pretreated with 3‐methyladenine (3‐MA, 5 mM) or wortmannin (WM, 100 nM) for 1 h, followed by 3 μM Pe‐C for the indicated times. (a) After treatment for 48 h, whole cell lysates were separated by SDS‐PAGE, then immunoblotting was carried out using anti‐LC3 antibody. (b) Cell viabilities were analyzed by MTT assay. (c) Sub‐G1 cell cycle phase was analyzed using flow cytometry. (d) Caspase‐3 activity was measured by a microplate reader at 405 nm.

Article Snippet: The activity of caspase‐3, ‐8, and ‐9 was measured using commercially available caspase colorimetric assay kits (Beyotime).

Techniques: SDS Page, Western Blot, MTT Assay, Flow Cytometry, Activity Assay