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. 2016 Sep 19;291(44):23237–23247. doi: 10.1074/jbc.M116.744508

FIGURE 1.

FIGURE 1.

Mice lacking miR-205 in thymic epithelial cells exhibit a more severe thymic stress response. A, Northern blots with a probe specific for miR-205 reveals its predominant expression in the thymus and skin versus the heart, kidney, liver, brain, spleen, and lung. U6 was used as a RNA loading control. B, real-time PCR experiments were used to evaluate the expression of miR-205 in sorted thymic epithelial cells (CD45−EpCAM+MHCII+) compared with isolated thymocytes (CD45+) and other stromal cells (CD45−EpCAM−MHCII−). C, real-time PCR revealed the expression of miR-205 in sorted thymic epithelial cells (CD45−EpCAM+MHCII+) before and following a systemic inflammatory response induced by poly(I:C) for 5 days. D, the elimination of miR-205 in thymic epithelial tissue was confirmed by Northern blots of RNA miR-205fl/fl:Foxn1-Cre mice compared with sibling controls. 5S RNA was used as a RNA loading control. E, histochemical analysis of the indicated thymii isolated from female mice 12 weeks of age was compared using H&E staining of 5-μm thymus sections (×10, scale bar 200 μm). F, thymus weight and total thymic cellularity was compared in the control and miR-205fl/fl:Foxn1-Cre female at 4, 8, and 12 weeks of age. No statistically significant differences were observed. G, histochemical analysis of the indicated thymii processed 2 days after a 2nd poly(I:C) injection was compared using H&E staining of 5-μm thymus sections (×10, scale bar 200 μm). H, thymus weight and total thymic cellularity were compared in the control and miR-205fl/fl:Foxn1-Cre mice at day 0 or 2 days following treatment of poly(I:C). I, the total number of cortical (EpCAM+MHCII+UEA1−BP-1+) and medullary (EpCAM+MHCII+UEA1+BP-1−) TECs was compared between the indicated mice, either untreated (day 0) or 2, 5, 8, and 11 days post-poly(I:C) injection (gating not shown). *, p < 0.04; **, p < 0.006; ***, p < 0.0002; ****, p < 0.00001 (Student's t test). Data are representative of mean ± S.E. from at least 3 mice per group. All mice used for this experiment were female.