Sp8 laser scanning confocal fluorescence microscope

Immunofluorescence staining was conducted as previously described (83 ). Cells were seeded on glass coverslips and cultured as indicated. Then, the cells were fixed in 4% paraformaldehyde in PBS for 30 min at 25 °C. After washing three times by PBS, the slides were blocked in the blocking buffer (5% goat serum, 2 mg/ml BSA, 0.1%Triton X-100 in PBS) for 1 h and incubated with primary antibodies overnight at 4 °C, followed by incubation with Alexa Fluor 488 goat anti-rabbit IgG or Alexa Fluor 594 goat anti-mouse IgG for 1 h at 25 °C. Subsequently, the slides were mounted with VECTASHIELD mounting medium containing DAPI and photographed with Leica SP8 laser scanning confocal fluorescence microscope.

Immunofluorescence staining was conducted as previously described (54 (link)). Cells were seeded on glass coverslips and transfected with indicated plasmids. Then, the cells were fixed in 4% paraformaldehyde in PBS for 30 min at 25 °C. After washing three times by PBS, the slides were blocked in the blocking buffer (5% goat serum, 2 mg/ml BSA, 0.1% Triton X-100 in PBS) for 1 h and incubated with primary antibodies overnight at 4 °C, followed by incubation with Alexa Fluor 488 goat anti-rabbit IgG for 1 h at 25 °C. Subsequently, the slides were mounted with VECTASHIELD mounting medium containing DAPI and photographed with Leica SP8 laser scanning confocal fluorescence microscope.