FIGURE 8.
IL23Rα R381Q and V362I were not stable. A, HEK293 cells stably expressing IL23Rα and IL23Rα variants were treated with cycloheximide (500 μm) for various times. Representative Western blot analysis of the cell lysate is shown here. B, IL23Rα mature (M) and immature (IM) levels over time under cycloheximide exposure are represented in percentages, which were calculated by taking IL23Rα levels at time 0 as 100%. One-phase exponential decay fit of the data was performed using GraphPad Prism 6.0, and the R2 value for the receptors are as follows: IL23Rα M (0.7212), IL23Rα IM (0.654), R381Q M (0.7460), R381Q IM (0.8905), G149R M (0.7655), G149R IM (0.876), V362I M (0.9559), V362I IM (0.9388). Error values were S.E. where n = 3. C. Half-life of IL23Rα and variants was calculated by using GraphPad Prism 6.0. Error values were S.D. where n = 3. Statistical significance calculated by ANOVA is denoted by the asterisk (*) compared with IL23Rα where p < 0.01; in addition, R381Q was significantly different from G149R and V362I with a p < 0.01, and G149R was significantly different from V362I with a p < 0.01.