Figure 6.

Effect of miR-124 inhibitor on phenotype of macrophages cocultured with neural and astroglial cells. (a) Flow cytometry analysis of the expression of CD45 (x axis) and MHC class II (y axis) in populations of CD11b⁺GFP⁺ gated BMDMs cultured alone (left contour plot) or cocultured with an astroglial (middle contour plot) or neuronal (right contour plot) cell line. Percentages of CD45^hiMHC class II⁺ cells are shown in upper right quadrants. BMDMs were isolated from ACTB-GFP transgenic mice that ubiquitously express GFP under the actin promoter. The cells were then analyzed for the expression of GFP, CD11b, MHC class II and CD45 using four-color flow cytometry. (b) qRT-PCR analysis of miR-124 expression in microglia, astroglial and neuronal lines, in BMDMs cultured alone or in CD11b⁺GFP⁺ BMDMs sorted from the cocultures. (c–e) BMDMs isolated from ACTB-GFP transgenic mice were cocultured with either an astroglial (c) or neuronal (d) cell line in the presence of anti–miR-124 or a control antagomir. (c,d) CD11b⁺GFP⁺ gated cells were analyzed for expression of CD45 (x axis) and MHC class II (y axis). Percentages of CD45^hiMHC class II⁺ cells are shown in upper right quadrants. (e) Data from three independent experiments are summarized, which show mean ± s.e.m. of percentages of CD45^hiMHC class II⁺ macrophages. *P < 0.05; **P < 0.01.