Figure 3.

miR-135a affects CD4⁺ T cell migration. T cell migration was determined using a modified 24-well Boyden Chamber. Genital tract tissue cell suspensions from mock or Cm infected (day 6) mice were added to the lower chamber (5 x 10⁵ per well), and transfected with miR-135a mimic or media alone for 24 hours prior to determination of migration. Enriched CD4⁺ T cells (from day 6 Cm infected splenocytes) were added to the upper chamber (5 x 10⁵ cell per well). Fourteen hours post addition of cells to the upper chamber, the lower chamber contents were used for (A) enumeration of migrated CXCR3-expressing CD4⁺ T cells by Flow cytometry, (B) comparison of relative CXCL10, CXCL12, and CCR5 gene expression in Cm infected cells transfected with and without miR-135a mimic to the uninfected/non-transfected control group (expression level normalized to 1), and (C) determination of IFN-γ secretion. Data shown are representative of 3 experiments. *p < 0.05 Student t test.