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. 2020 May 29;10(4):673–697. doi: 10.1016/j.jcmgh.2020.05.007

Figure 9.

Figure 9

JAK2, TYK2, and STAT3 are required for IL23-induced antimicrobial pathways and intracellular bacterial clearance. (A) MDMs were treated with 10 ng/mL IL23 or IL12 for 60 minutes and assessed for fold increase in phospho-STAT3 by flow cytometry (n = 4). (B–J) MDMs were transfected with scrambled or the indicated siRNAs, alone or in combination (comb). (B) STAT3 expression by flow cytometry (mean fluorescence intensity [MFI]) (n = 4). (C) Cell death per annexin V+ cells (n = 4). A total of 50–100 J/m2 UV-treated cells served as a positive control. (D–J) MDMs then were treated with (D–F and J) IL23 or (G–I and J) IL12 for 48 hours. (D–I) ROS production or expression of the indicated molecules by flow cytometry (MFI) (n = 8 from 2 independent experiments for ROS, p40phox, NOS2, LC3II, ATG16L1; n = 4 for p47phox, p67phox, ATG5 with similar results in an additional 4 donors). (J) Intracellular bacterial clearance (CFU) (n = 4). Means + SEM. ∗∗P < .01, ∗∗∗P < .001, †P < 1 × 10-4, and ††P < 1 × 10-5. NT, no treatment; scr, scrambled; Tx, treatment.