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gp38+CD31+ LN stromal cells express and present Tyr369 in an Aire-independent manner. (a) Tyrosinase (Tyr) and β-actin expression in CD45neg and CD45+ LN populations from B6 and albino animals was determined using 40-cycle RT-PCR. Data shown are representative of four independent experiments. (b) Proliferation of CFSE-labeled naive Thy1.2+ FH T cells co-cultured with Thy1.1+ CD45neg (black bars) or CD45+ (white bars) cells purified from the LNs of tyrosinase+ and albino animals was assessed after 86 h. Left, representative experiment; right, summary data for two independent experiments. (c) LN stromal cell populations defined based on expression of gp38 and CD31. The representative plot is gated on CD45neg cells. Numbers indicate percentage of total CD45neg cells. (d) Tyrosinase and β-actin expression in CD45+ cells and four LN stromal cell subpopulations identified by differential expression of gp38 and CD31. Purified populations were obtained by electronic cell sorting and 40-cycle RT-PCR was performed. Data shown are representative of six independent experiments. (e) Proliferation of CFSE-labeled naive Thy1.2+ FH T cells co-cultured with Thy1.1+ cells of the indicated purified LN stromal cell subpopulations from tyrosinase+ (black bars) and albino (white bars) animals was assessed after 86 h. Left, representative experiment; right, summary data for three independent experiments. Error bars indicate SEM.