Figure 3.

HOPX and SOX2 are specifically expressed in human Th17 cells

Naive CD4⁺ T cells cultured with anti-CD3/anti-CD28 alone (Th0) or with the addition of TGF-β, IL-6, IL-1β, and IL-23 (infl. Th17) or in the absence of IL-1β (reg. Th17) were analyzed by RNA sequencing. The Venn's diagram between genes upregulated in inflammatory Th17 versus regulatory Th17 at 48 h and 5 days unveils 18 genes regulated at both time points (group X) and 421 genes specifically upregulated at 5 days (group Y) (A). All the annotated genes included in group X and 10 of 421 genes in group Y are listed in panel (B) HOPX and SOX2 expression obtained by RNA sequencing (reported as DEseq2 normalized count) (n = 5) and by q-PCR (n = 4) (C) at 48 h and 5 days and by Western blot (n = 5) at 5 days (D) were analyzed in Th0, inflammatory Th17 (TGF-β, IL-6, IL-1β and IL-23), and regulatory Th17 (TGF-β, IL-6, and IL-23) cells. Expression of HOPX transcript (n = 5) (E) and protein (n = 4) (F), SOX2 transcript (n = 6) (G) and protein (n = 4) (H) in Th0, Th1, Th2, Th17, and Treg profiles differentiated in vitro from naive CD4⁺ T cells of HD. Data are represented as mean ± SEM (For pairwise conditions: Student's T-tests; For three or more conditions: One-way ANOVA test; ∗p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001). Th1: IL-12; Th2: IL-4; Th17: IL-1β, IL-6, TGF-β, and IL-23; iTreg: IL-2 and TGF-β; Th0: no cytokines. Inflammatory Th17 = infl. Th17; regulatory Th17 = reg. Th17.