Prolactin Increases the Frequency of Follicular T Helper Cells with Enhanced IL21 Secretion and OX40 Expression in Lupus-Prone MRL/lpr Mice

Figure 5

Activation and signaling of differentiated T_FH cells in vitro in the presence of PRL. T_naïve cells from 9-week-old C57BL/6 and MRL/lpr mice were purified by MACS and differentiated to T_FH with or without PRL. (a) Expression of OX40 (MFI) and representative histograms of OX40 expression in T_FH cells from MRL/lpr mice. (b) Percentage and representative Zebra plots of T_FH OX40⁺ cells. (c) Expression of IL21 (MFI) and representative histogram of IL21 expression in T_FH cells from MRL/lpr mice. (d) Percentage and Zebra plots of T_FH IL21⁺ cells. (e, f) T_naïve and T_FH cells were preincubated for 30 min with the inhibitor of STAT3 (Stattic). For STAT3 inhibition, T_naïve cells were differentiated to T_FH, left to rest for 8 h, and then incubated for 30 min with PRL to subsequently determine the MFI of pSTAT3 (histogram). For the plot, pSTAT3 is reported as fold change taken as baseline levels found in T_naïve or T_FH cells only treated with medium. (f) Percentage and Zebra plots of pSTAT3. Six different experiments were performed; each experiment was done in duplicate. Pooled data are presented as the ; and using ANOVA.

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