As 1109

Primary mouse chondrocytes were rinsed in PBS and fixed with 4% paraformaldehyde for 15 min at room temperature. After being permeabilized with 0.3% Triton X-100 for 5 min, the cells were blocked with 1% BSA for 30 min and incubated with mouse anti-MMP-13 (1:300, Abcam, ab219620) and anti-Col2a1 (1:500, Abcam, ab34712) antibodies at 4 °C overnight. The samples were then washed with PBS (4 °C) three times and incubated with a Cy3-conjugated goat anti-rabbit secondary antibody (1:50, ASPEN, AS1109) at 37 °C for 30 min. TUNEL staining of chondrocytes was performed with a TUNEL Assay Kit (Roche, Basel, Switzerland) according to the manufacturer’s protocol.

Bladder cancer cells (1 × 10⁶) were plated in 6-well chamber slides and exposed to blue laser for 4 J/cm², 8 J/cm², or no irradiation. After 48 h, the cells were fixed with 4% paraformaldehyde for 30 min and incubated with blocking solution for 10 min at 20°C for permeabilization and non-specific antigen blocking. Next, the cells were incubated with rabbit anti-Ki67 primary antibody (1:150 dilution, 27309-1-AP, Proteintech, USA) overnight at 4°C. The cells were then incubated with goat anti-rabbit antibody for 50 min (AS-1109, ASPEN Biotechnology, China) as a secondary antibody, and the nuclei were stained with 4′,6-diamidino-2-phenylindole (AS1075, ASPEN Biotechnology, China). Images of five random fields were obtained for each group using a fluorescence microscope (BX51, Olympus, Japan). Ki67-positive cells were stained red in the nucleus. Five random fields were chosen from each slice, and the percentage of Ki67-positive cells was determined.