The responses of <i>Usp21</i>^-/- (-/-) and control wild type (+/+) bone marrow derived macrophages and dendritic cells to TLR and TNFR stimulation.

<p>(A-B)The cells were stimulated over 18 hours with LPS at 10 and 100ng/ml, poly(IC) at 5 and 25 μg/ml, or TNFα at either 5 or 10ng/ml. (A) Production of cytokine TNFα was assessed by intracellular staining and flow cytometry, with the addition of Brefeldin A at 3μg/ml at 2 hours following the beginning of the stimulation. (B) Production of cytokines IL-6 and TNFα was assessed by ELISA. (C) Bone marrow derived macrophages from <i>Usp21</i>^-/- (-/-, dashed line) and control wild type (+/+, solid line) mice were stimulated over 1.5 hours with LPS 100ng/ml, poly(IC) 25 μg/ml, or TNFα 10ng/ml, and analyzed by flow cytometry for IκBα levels and p65 NFκB S536 phosphorylation; data is presented as fold change in MFI over the untreated sample. Bars represent means ± SEM, datasets are from 4–5 mice per group with the cells from each mouse stimulated and analyzed in duplicate; MFI-mean fluorescence intensity; statistical analysis by ANOVA with Bonferroni post-hoc test, ** p<0.01, * p<0.05, and non-significant if no p-value is provided. All datasets were acquired however only datasets with a significant difference between stimulated and unstimulated samples are presented.