Figure 1.

Early, persistent, and late genes acquired during human Th17 cell polarization reflect specific functions

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 (Th17) (n = 5) were analyzed by q-PCR for the expression of RORC transcript (A). RNA sequencing of Th17 and Th0 cells (n = 5) at 48 h and 5 days of differentiation (B): number of genes differentially modulated in Th17 versus Th0 at each time point (C); Venn's diagram shows the number of genes specifically modulated (either upregulated or downregulated) in Th17 profile (Th0 versus Th17) at 48 h (early) and 5 days (late) and persistently modulated (48 h Th17 versus 5 days Th17) (D); expression of selected Th genes included in A, B, and C modules reported as DEseq2 normalized count (E); bar graph representation of gene ontology for genes in A, B, and C modules (F); expression of selected Th17 transcription factors (G and H). Each replicate, including Th0_48h, Th17_48h, Th0_5d, Th17_5d, was performed in the same donor (paired cultures). Data are represented as mean ± SEM (Student's T-tests; ∗p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001; ∗∗∗∗p ≤ 0.0001).