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.