4 6 diamidino 2 phenylindole dihydrochloride dapi solution

PC9 cells were fixed in 4 wt% paraformaldehyde (Sigma, St Louis, MO, USA) for 30 min and rinsed three times with PBS for 10 min each time. The samples were immersed in 0.2% Triton X-100 for 10 min, rinsed three times for 10 min each time with PBS, and then blocked in 4% goat serum for 1 h at room temperature. Then sample sections were incubated overnight in anti-E-cadherin rabbit monoclonal antibody (1:200, Cell Signaling Technology) and anti-vimentin rabbit monoclonal antibody (1:200, Cell Signaling Technology) at 4°C. After rinsed with PBS three times, the samples were subsequently incubated in anti-rabbit immunoglobulin G secondary antibody conjugated with fluorescein isothiocyanate (FITC) or APC (1:100, eBioscience) for 1 h in the dark. For nuclei observation, the samples were dipped in 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) solution (Sigma, 3 μg/mL) and immediately rinsed with PBS. In the stained image, the E-cadherin displayed red fluorescence, the vimentin displayed green florescence, and the nuclei displayed blue fluorescence.

MM.1S cells treated for 24h were fixed, blocked and incubated overnight with a primary anti-α-tubulin antibody (Sigma) and counterstained with Alexa Fluor 488 conjugated anti-mouse immunoglobulin G (IgG) secondary antibody (Life Technologies, Waltham, MA, USA) and a 4′,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI) solution (Sigma).