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. 2020 Dec 4;11:588543. doi: 10.3389/fimmu.2020.588543

Figure 3.

Figure 3

ABT 317 does not affect GM-CSF signaling or blood cell counts. (A) FACS gating process of granulocytes is shown. (B–E) Peripheral blood was cultured with or without 2 µM ABT 317 or 1 µM JAK1/JAK2 inhibitor for 1 h prior to the addition of GM-CSF for 30 min to analyze the mean fluorescence intensity (MFI) expression of phosphorylated STAT5 in granulocytes (B, C) and the percentage of pSTAT5 positive granulocytes (D, E). Data show individual mice (n = 3) and mean ± SEM for three independent experiments. (D) Representative plots are shown. (F) Peripheral blood was collected from 6–8-week-old NOD mice treated with vehicle, 10 mg/kg ABT 317 or 3 mg/kg JAK1/JAK2 inhibitor for 3 days and stimulated ex vivo with GM-CSF. The MFI of phosphorylated STAT5 in granulocytes is shown for n = 5–7 individual mice from two independent experiments. (G, H) Peripheral blood was collected from NOD mice treated with vehicle, 10 mg/kg ABT 317 or 3 mg/kg JAK1/JAK2 inhibitor for 3 weeks. (G) White blood cell counts and (H) hemoglobin levels are shown for n = 7–10 individual mice from three independent experiments. Data are shown as representative plots and the mean ± SEM of pooled data. Statistical significance: *P = 0.0365, ***P = 0.0004, one-way ANOVA with Tukey’s test for multiple comparisons.