Fig. 7: Inhibition of TNFα blocks increase in number of TH+ monocytes and amount of TH per monocyte.

a and b Acutely isolated monocytes from three healthy donors were seeded at 2 million cells per well in duplicate ultra-low-adherence plates, and treated with TNFα (17 ng/mL), XPro1595 (50 ng/mL), IL6 (17 ng/mL) or combinations thereof as indicated. c In samples assayed by flow cytometry, using counting beads as a reference value to quantify the number of TH+ cells, co-incubation with TNFα and XPro1595 significantly reduced the number of TH+ monocytes relative to TNFα treatment alone. Treatment with IL6 or IL6 + XPro1595 resulted in no significant change in the number of TH+ monocytes. Values are represented as fold change relative to vehicle (n = 3 per group, one way ANOVA, *p < 0.05, **p < 0.01). d TH concentration (picogram per milligram total protein) significantly increases upon TNFα treatment, and is reduced significantly to baseline levels following co-incubation with TNFα and XPro1595. Neither IL6 nor IL6 + XPro1595 significantly increased TH quantity (n = 3 per group, one way ANOVA, *p < 0.05). Data are shown as ±SEM.