Fig. 5

Retroviral transduction of Bhlhe40 restores the pathogenic function of Satb1-deficient Th17 cells. a qPCR of Bhlhe40 mRNA expression in CD25⁻ CD44^low CD4⁺, CD25⁻ CD44^high CD4⁺, PPs eYFP⁺ CD4⁺, and EAE LNs eYFP⁺ CD4⁺ T cells. EAE LN eYFP⁺ Th17 cells were sorted on days 7–14 after EAE induction. b qPCR of Bhlhe40 mRNA expression 3 days after Bhlhe40 was retrovirally transduced in naive CD25⁻CD44^low CD4⁺ T cells. c Mean (+s.e.m.) clinical scores on the days after EAE induction in Rag2^−/− mice; the following conditions were followed for the transfer of NGFR⁺ CD4⁺ T cells (2 × 10⁶): (1) control CD4⁺ T cells transduced with the Bhlhe40 vector (n = 5), (2) control CD4⁺ T cells transduced with the control vector (n = 5), (3) Th17^Satb1KO CD4⁺ T cells transduced with the Bhlhe40 vector (n = 6), and (4) Th17^Satb1KO CD4⁺ T cells transduced with the control vector (n = 5). The incidence of EAE was as follows: (1) 5/5, (2) 5/5, (3) 6/6, and (4) 2/5. ***P < 0.0001 (two-way ANOVA with Bonferroni’s post-test). d, e Flow cytometry of CD4⁺ T cells (top) and NGFR⁺ eYFP⁺ CD4⁺ T cells (bottom) in draining LNs (d) and spinal cord (e) as in (c) for IL-17 and GM-CSF expression on day 14 after EAE inductions. The frequencies of GM-CSF⁺ in eYFP⁺ CD4⁺ T cells are shown. The bar graphs (a, b, d, e) show the mean ± s.d. (n = 3). The results are representative of two independent experiments. **P < 0.001; ***P < 0.0001 (two-tailed unpaired Student’s t-test or one-way ANOVA)