Figure 5.

Ara-LAM downregulated foxp3 gene expression by IRF1 mediated suppression in Tregs during L. donovani infection. (A) CD4⁺CD25⁻ T cells and CD4⁺CD25⁺ T cells were purified from spleen of differently treated mice by MACS (section ‘Materials and Methods’) and collected in TRIZOL for mRNA extraction. The expression of IRF1, IRF4 and IRF8 was analyzed by semiquantitative RT-PCR. The data are representative of three independent experiments yielding similar results. (B–D) CD4⁺CD25⁺Tregs, from spleen of differently treated mice, were purified as described earlier. IRF1 binding at Foxp3 promoter (IP:IRF1), SMAD3 binding at Foxp3 promoter (IP:SMAD3) and histone 3 acetylation at the Foxp3 promoter were analyzed by ChIP assay. Data represent means ± SD for three sets of experiments.