Cells were incubated in 24-well plates under standard cell culture conditions (5 × 103 cells per well). After 12 h, cells were blocked with 5% BSA for 15 min, stained with B7-H3 antibody (Abcam, ab227679) for 1 h, Cy3-conjugated secondary antibody (Beyotime, A0516) for 40 min and DAPI (Beyotime) in the dark. Images were captured on a fluorescence microscopy.
Tumor tissues from the T cell group mice were collected and immediately froze at −80°C. Sections were fixed in pre-chilled acetone-methanol (1:1) for 20 min at −20°C and then allowed to air-dry for 10 min before being blocked with 5% BSA for 30 min. Subsequently, sections were stained with B7-H3 antibody (Abcam, ab227679) for 1 h, FITC-conjugated secondary antibody (Beyotime, A0562) for 40 min and DAPI (Beyotime) in the dark. Images were captured on a fluorescence microscopy.
Tumor tissues from the T cell group mice were collected and immediately froze at −80°C. Sections were fixed in pre-chilled acetone-methanol (1:1) for 20 min at −20°C and then allowed to air-dry for 10 min before being blocked with 5% BSA for 30 min. Subsequently, sections were stained with B7-H3 antibody (Abcam, ab227679) for 1 h, FITC-conjugated secondary antibody (Beyotime, A0562) for 40 min and DAPI (Beyotime) in the dark. Images were captured on a fluorescence microscopy.