Figure 6.

Effect of isoflurane on activating mutants. A) Surface expression and ICAM-1 binding to α-I306A and β-L132A mutants. LFA-1 surface expression was probed by TS2/4 antibody. Data are shown as means ± se of 3 independent experiments of triplicates. MFI, mean fluorescence intensity. B) ICAM-1 binding of mutants normalized with surface expression. Normalized MFI of ICAM-1 binding was calculated as [MFI of ICAM-1 binding of mutant] × {[(MFI of surface expression of WT) − (MFI of surface expression of mock transfectant)]/[(MFI of surface expression of mutant) − (MFI of surface expression of mock transfectant)]}. One-way ANOVA with Tukey post hoc test was performed to compare data within Mg²⁺/Ca²⁺ or Mn²⁺ group. *P < 0.05 vs. WT Mg²⁺/Ca²⁺. C) ICAM-1 binding was performed at 1 mM Mn²⁺ with or without 2% isoflurane. Relative binding (%) was defined as [(MFI of ICAM-1 binding with 2% isoflurane) − (MFI of mock transfected sample)]/[(MFI of ICAM-1 binding without isoflurane) − (MFI of mock transfected sample)] × 100 (%). Samples that did not get exposed to isoflurane were considered 100% binding (control sample). Data are shown as means ± se of 3 independent experiments, each with triplicates. Statistical analysis was done using 1-way ANOVA with Tukey post hoc analysis. *P < 0.05 vs. control.