SHIP1 mediates IL-10 inhibition of miR-155.

<p>(A) Perimacs were extracted from either WT or SHIP1 KO mice, and stimulated with 10 ng/ml LPS or LPS +10 ng/ml IL-10. miR-155 expression levels were measured by real time PCR and plotted relative to the LPS alone sample in each cell type. (B) SCRMB and SHIP1 siRNA transduced cells were treated with 2 µg/ml Dox for 48 hours or left untreated prior to immunoblotting analysis for SHIP1 and STAT3 (loading control). Band intensities were quantified using the Quantity One Software. Statistical significance between treatments were calculated by a two-way ANOVA test with a 95% confidence (****<i>p</i><0.0001). (C) SCRMB and SHIP1 siRNA transduced cells were treated with 2 µg/ml Dox for 48 hours and then stimulated with LPS +/− IL-10 for 2 or 4 hours. Expression levels of pri-miR-155 in the 2-hour samples and miR-155 in the 4-hour samples were measured by real time PCR and plotted relative to the LPS alone samples. (D) RAW264.7 cells were left untreated, treated with 10 µM AQX-MN100 or ethanol control for 30 minutes before being stimulated with LPS or LPS+AQX-MN100 for 4 hours. Expression level of miR-155 was measured by real time PCR and plotted relative to LPS samples. (E) AKT-ER transduced cells were treated with 150 nM 4-HT for 20 minutes or left untreated before being stimulated by LPS +/− IL-10 for 4 hours. Expression level of miR-155 was measured by real time PCR and plotted relative to the LPS alone samples. Statistical significance between stimulation conditions was calculated by a two-way ANOVA test with a 95% confidence (**<i>p</i><0.01, ***, <i>p</i><0.001, ****<i>p</i><0.0001). Results were observed in at least two independent experiments.