AntagomiR to miR-223 reduces macrophage differentiation and cell survival. In 6-well plates, THP-1 cells (0.5 × 106) and monocytes (1.0 × 106) were plated in 2.4 mL of media then transfected with miR-223 antagomiR and incubated with PMA or GM-CSF, respectively, in X-VIVO 15 medium. Both nonadherent and adherent cells were collected and analyzed for cellular differentiation and survival. Cellular adherence was measured by crystal violet assay in the antagomiR-transfected (A) THP-1 cells and (B) monocytes. Shown is the average absorbance for crystal violet ± SEM (n = 3 in duplicate). AntagomiR-transfected (C) THP-1 cells and (D) monocytes were analyzed for gene expression as indicated. The average relative copy number is shown ± SEM from 3 independent experiments. Caspase-3 assay was performed for (E) THP-1 cells (n = 3) and (F) monocytes (n = 5) transfected with the miR-223 antagomiR then treated with PMA or GM-CSF, respectively. Shown is the average fold change ± SEM compared with untransfected cells.
