Anti gli1

Cells were immersed in PBS and seeded in glass slides at a concentration of 1 × 10⁴/ml. The slides were fixed in 4% paraformaldehyde for 15 min, immersed in PBS 3 times for 3 min each time and subsequently immersed in 0.5% Triton X‐100 in PBS at room temperature for 20 min. Next, the cells were blocked with 5% skim milk powder in PBST for 30 min at room temperature, and the following diluted primary antibodies were added: rabbit anti‐mouse and anti‐Gli1 (R&D, USA). The antibodies used were rabbit anti‐human and were diluted at a concentration of 1:200. The cells were incubated overnight at 4°C and subsequently incubated with a fluorescent secondary antibody (1:500, goat anti‐rabbit antibody, CST, USA) for 1 h at room temperature. An appropriate amount of DAPI (1:5000, Thermo, USA) was added. The cells were incubated at room temperature for 5 min in the dark, and the nucleation status of Gli1 in each treatment group was observed under an inverted fluorescence microscope (Nikon, Japan).