IL-12 and IL-23 induce naïve CD4⁺ T cells to express multiple Tfh-associated molecules. Human naïve CD4⁺ T cells were stimulated with CD3-CD28 mAb-coated beads in the presence of the indicated cytokines to analyze expression of Tfh-associated molecules. (A-C) Expression of IL-21 (after 6-hour restimulation with PMA and ionomycin), CXCR5, and ICOS on day 3. (A) A representative result of IL-21 expression. (B) A representative result of CXCR5 and ICOS expression. (C) Each dot represents data obtained from different donors. Mean ± standard deviation (SD); n = 3-13. One-way ANOVA: *P < .05; **P < .01; and ***P < .001. (D) Expression of BCL6, PRDM1, BATF, and MAF transcripts of CD4⁺ T cells cultured for 2 days. Abundance of transcripts was measured by Nanostring. Normalized to the abundance in the control conditions following gene normalization with 7 housekeeping genes. Mean ± SD; n = 3. One-way ANOVA. (E) Analysis of Bcl-6 expression by flow cytometry. CD4⁺ T cells were primed for 3 days in the presence of indicated cytokines. Mean ± SD; n = 3. One-way ANOVA.