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. 2012 Feb 13;209(2):243–250. doi: 10.1084/jem.20111174

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© 2012 Johnston et al.

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Figure 2. — Lack of STAT5 signaling enhances T_FH cell differentiation. STAT5^fl/fl SM cells were transduced with an RV expressing Cre recombinase (Cre) or were not transduced but treated similarly (control). (A) Representative histograms of phospho-STAT5 levels in SM cells, with and without IL-2 stimulation. The percentage of cells that was phospho-STAT5⁺ is indicated. (B–J) Cre⁺ and control SM cells were adoptively transferred into C57BL/6J mice that were subsequently infected with LCMV. Splenocytes were analyzed 8 (B–F) or 4 (G–J) d after infection. Data are a composite of two independent experiments, and n = 8/group. (B) Quantitation of SM cells as a percentage of all CD4⁺ T cells. (C) Representative histograms gated on SM cells or on CD4⁺ T cells from an uninfected C57BL/6J mouse (naive). (D) Representative FACS plots gated on SM cells, with T_FH cells (SLAM^low CXCR5^high) boxed. Quantitation of SM T_FH cells as a percentage of total SM cells. ***, P < 0.0001. (E) Representative FACS plots gated on SM cells, with GC T_FH cells (CXCR5^high GL7^high) boxed. Quantitation of SM GC T_FH cells as a percentage of total SM cells. **, P = 0.01. (F) IL-21 production by SM cells after PMA/ionomycin stimulation in vitro. Quantitation of IL-21⁺ SM cells as a percentage of total SM cells. (G) Quantitation of SM cells as a percentage of total CD4⁺ T cells. (H) Representative histograms of Bcl6 expression, gated on SM cells or on CD4⁺ T cells from an uninfected C57BL/6J mouse (naive). Quantitation of SM cell Bcl6 MFI. **, P = 0.0012. (I) Representative FACS plots gated on SM cells, with T_FH (Bcl6^high CXCR5^high) boxed. Quantitation of SM T_FH cells as a percentage of all SM cells. ***, P < 0.0001. (J) Representative FACS plots gated on SM cells, with non-T_FH cells (CD25^high CXCR5^low) boxed. Quantitation of non-T_FH SM cells as a percentage of total SM cells. ***, P < 0.0001. Error bars depict the standard error of the mean.