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Figure 2. Proteolysis of ICAM-1 on KB cells by Rgp. (A) KB cells were fixed with 3% paraformaldehyde for 3 min at 4°C. After being washed with PBS, unfixed or fixed cells were incubated with 0.3 µmol/L HRgpA for 1 hr at 37°C, and then expression of ICAM-1 on KB cells was assessed by flow cytometry. Representative findings of three independent experiments are expressed as the mean ± SD of the MFI. **P < 0.01 vs. respective control. (B) Purified cell membrane of KB cells was untreated or treated with 0.3 µmol/L RgpB for the period indicated at 37°C, and then the expression of ICAM-1 was analyzed by Western blotting with anti-ICAM-1polyclonal Ab. (C) Purified cell membrane of KB was untreated (lane 1) or treated with 0.3 µmol/L RgpB for 1 hr without (lane 2) or with 3 µmol/L FPR-cmk (lane 3), and 3 µmol/L Z-FK-cmk (lane 4) pre-treatments for 10 min at room temperature. Then, the expression of ICAM-1 was analyzed by Western blotting with anti-ICAM-1polyclonal Ab. Molecular-mass markers (kDa) are shown on the left. Findings are representative of three independent experiments.
