Cytoskeleton changes were detected by F‐actin staining. T24 cells (5 × 104 cells/well) were seeded into 12‐well plates covered with cell slides made of high‐quality glass (WHB scientific) and treated with MH for 24 h. After fixation with 4% paraformaldehyde, cell slides were stained with FITC‐conjugated phalloidin (Yeasen) for 30 min and DAPI (Vector) for 10 min at room temperature. For F‐actin staining, tumors were frozen in OCT (Servicebio) embedding medium, and 20 μm sections were cut sequentially and mounted on superfrost plus slides. Slices were permeabilized in 3% PBST and stained with FITC‐conjugated phalloidin for 30 min and DAPI for 10 min at room temperature. Images were acquired with a laser confocal microscope (Model IX83, Olympus).
SEM was performed with a Hitachi Regulus SU8100 field emission scope using an accelerating voltage of 3 kV. T24 cells were seeded in 6‐well plates (1 × 105 cells/well) and fixed with an electron microscope fixative (Servicebio) for 24 h at 4°C. Cell morphology was observed and representative pictures were acquired during SEM imaging.
SEM was performed with a Hitachi Regulus SU8100 field emission scope using an accelerating voltage of 3 kV. T24 cells were seeded in 6‐well plates (1 × 105 cells/well) and fixed with an electron microscope fixative (Servicebio) for 24 h at 4°C. Cell morphology was observed and representative pictures were acquired during SEM imaging.