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Figure 3: ERα promotes CD4⁺ T regulatory cell response. Spleen cells from C57Bl/6, ERαKO and ERβKO mice infected 7 days earlier with 10² PFU CVB3 were labeled with antibodies to CD4, CD1d and CD25, fixed, permeabilized and labeled with antibody to FoxP3 for flow cytometry analysis. (A) Cells were gated on the CD4⁺ cell population from an infected C57Bl/6 mouse, and then evaluated for CD25 and FoxP3 expression. The CD4⁺CD25^-FoxP3^- and CD4⁺CD25⁺FoxP3⁺ subpopulations were evaluated for CD1d expression. The line in the CD1d graph at far right indicates mean intensity fluorescence for the CD4⁺CD25^-FoxP3^- population and indicates that the CD4⁺CD25⁺FoxP3⁺ cells express higher levels of CD1d. (B) Summary of percentage of CD4⁺ cells which are CD25⁺FoxP3⁺ for 5-6 mice/group (mean ± SEM). (C) Summary of mean fluorescence intensity of staining for CD4⁺ cells which are either CD25^-FoxP3^- or CD25⁺FoxP3⁺ for 5-6 mice/group (mean ± SEM). *Significantly different than C57Bl/6 mice at p<0.05.