Autophagosomes were not co-localized with lysosomes in physalin B-treated cells. (A) Autophagosomes were not co-localized with lysosomes. GFP-LC3/RFP-LAMP1-co-transfected cells were treated with 10 μmol/L physalin B, 0.1% DMSO (negative control) and 5 μmol/L rapamycin (positive control) for 12 h. Localization of GFP-LC3 and RFP-LAMP1 were observed under an Olympus Fluoview FV1000 confocal microscope (Olympus, Tokyo, Japan). (B) Microtubule cytoskeleton was changed in physalin B-treated cells. Cells were treated with 10 μmol/L physalin B, 0.1% DMSO (negative control) and 100 ng/mL nocodazole (positive control) for 12 h. Samples were then prepared as mentioned in the “Materials and methods” section, and the status of microtubules was observed using an Olympus confocal microscope (Olympus, Tokyo, Japan). (C) F-actin microfilaments were changed in physalin B-treated cells. Cells were treated with 10 μmol/L physalin B, 0.1% DMSO (negative control) and 5 μmol/L rapamycin for 12 h. Samples were then prepared as mentioned in the “Materials and methods” section, and the status of F-actin microfilaments was observed using an Olympus confocal microscope (Olympus, Tokyo, Japan).