Fitc conjugated secondary goat anti rabbit antibodies

The immunofluorescence was used to verify the expression location of RNPC1a and ERα. Briefly, the breast cancer cells were plated in 24-well plate at a density of 5×10⁴ cells/well and incubated for 12h. After washed with phosphate-buffered saline (PBS, pH=7.4) twice, the cells were fixed with paraformaldehyde for 20 min and penetrated by 0.5% Tritonx-100 for 10 min, followed by blocking for 1 h in blocking buffer. Then cells were incubated with primary antibody overnight at 4 °C. After washed with PBS three times, cells were incubated for 1 h in the dark with FITC-conjugated secondary goat anti-rabbit antibodies (Invitrogen, USA). The cells were then washed and stained with 4, 6-diamidino-2-phenylindole (DAPI) for 5 min. Immunostaining was observed under a Zeiss fluorescence microscope at 400× magnification.

The expression location of RNPC1a and PR was conducted by the immunofluorescence, as previously described [37 (link)]. Briefly, the breast cancer cells were plated in 24-well plate at the density of 5×10⁴ cells per well. After 36 h incubation, the cells were washed with phosphate-buffered saline (PBS, pH 7.4) twice, and then fixed with paraformaldehyde for 20 min and penetrated by 0.5% Tritonx-100 for 10 min, followed by blocking for 1 h in blocking buffer. Then the cells were incubated with primary antibody at 4°C overnight. After washed with PBS three times, the cells were incubated for 1 h in the dark with FITC-conjugated secondary goat anti-rabbit antibodies (Invitrogen, USA). The cells were then washed and stained with 4, 6-diamidino-2-phenylindole (DAPI) for 5 min. Immunostaining was observed under a Zeiss fluorescence microscope at 400× magnification.