Anti rbpj

Cells were inoculated on sterile crawlers placed in 24-well plates and performed as described previously when 80% confluence was reached [47 (link)]. Cells were incubated with anti-RBPJ (1:200, Beyotime Biotechnology, Shanghai, China) overnight at 4 °C. The cells were washed and incubated with the fluorescent secondary antibody for 1 h at RT, then washed with PBST followed by DAPI staining for 5 min. After processing, the images were analyzed using an LSM 710 confocal laser microscope device (Zeiss, Oberkochen, Germany).

After treatment and detection of total protein concentration with the BCA Protein Assay Kit (Epizyme Bio, Shanghai, China), total proteins were collected from cells for protein blotting experiments [22 (link),46 (link)]. Anti-Notch1 (1:300, Wanlei Bio, Shenyang, China), anti-p-Notch1 (1:300, Wanlei Bio, China), anti-Notch2 (1:300, Wanlei Bio, China), anti-RBPJ (1:1000, Beyotime Biotechnology, Shanghai, China), anti-Runx2 (1:300, Wanlei Bio, China), anti-Col2a1 (1:1000, Beyotime Biotechnology, Shanghai, China), anti-BMP2 (1:300, Wanlei Bio, China), anti-MMP9 (1:300, Wanlei Bio, China), anti-MMP13 (1:300, Wanlei Bio, China), anti-GAPDH (1:1000, Proteintech, Wuhan, China) primary antibodies were used.