Immunofluorescence for Cell Cytoskeletal
Immunofluorescence for cell cytoskeletal rearrangement was performed using Glass Bottom Cell Culture Dish (801001; Nest, Wuxi, China) with Laser scanning confocal microscope (leica, TCS SP8, Shanghai, China). Before the experiment, TC was pretreated with different concentrations (0, 6, 12 μg/mL) for 48 h. Then 2.5 × 104 of AMDC was resuspended in 2 mL complete DMEM containing 10% UY at Glass Bottom Cell Culture Dish. Cells were then incubated at 37 °C in a 5% CO2 for 24 h with 50% AMDC. In brief, cells were fixed with 4% paraformaldehyde for 10 min at Room temperature, followed by 0.5% Triton X-100 for 5 min, 150 nM of TRITC-Phalloidin (Solarbio, CA1610, Beijing, China) with 1% BSA (Albumin Bovine V) (Solarbio, A8020, Beijing, China) in PBS for 30 min avoid light, DAPI 5 min. Before each next step, washing with PBS 3 times for 5 min. Then under the Laser scanning confocal microscope.