Panel A contains four bar graphs. The first bar graph shows the percentage of mCherry-positive HEK cells for various interleukin-23 receptor (IL23R) variants. The horizontal axis lists the variants, and the vertical axis shows the percentage of mCherry-positive cells. The second bar graph shows the percentage of interleukin-12 receptor subunit beta-1-positive cells within the mCherry-positive population. The horizontal axis lists the variants, and the vertical axis shows the percentage of interleukin-12 receptor subunit beta-1-positive cells. The third bar graph shows the levels of interleukin-23 receptor messenger RNA. The horizontal axis lists the variants, and the vertical axis shows the base-10 logarithm of 2 raised to the power of negative delta cycle threshold. The fourth bar graph shows the levels of interleukin-12 receptor subunit beta-1 messenger RNA. The horizontal axis lists the variants, and the vertical axis shows the base-10 logarithm of 2 raised to the power of negative delta cycle threshold. Panel B contains multiple western blots from HEK cells expressing different interleukin-23 receptor alleles and stimulated with interleukin-23 or interferon alpha-2. The horizontal labels identify the receptor variants, including wild type and multiple mutant forms.
Analysis of IL23R variants from the general population. (A) Frequency of mCherry+ and of IL-12Rβ1+ cells in the mCherry+ population and levels of IL23R and IL12RB1 mRNA in HEK cells transiently transduced with plasmids encoding IL-12Rβ1 and the various IL-23R variants expressed together with the mCherry reporter gene and a luciferase reporter. Order of plasmids is as follow: NT, EV, followed by IL23R WT, C115Y, G300V, R381Q, Q3H, A55T, R86Q, G149R, K150T, V159M, V160A, L193F, A199V, S221F, P306S, L30P, V362I, G300V+V362I, L372F, I373F, Q487H, and S559R. Bars indicate the mean ± SD for three independent biological replicates. (B) Detection by western blotting of pSTAT3 after IL-23 (10 ng/ml) and IFN-α (1 ng/ml) stimulation in HEK cells transfected with the indicated hypomorphic IL23R alleles, with total STAT3 and vinculin as loading controls. At least two independent experiments for each variant were performed. The blot is duplicated in Fig. 1 B. Source data are available for this figure: SourceData FS1.