(B) Representative plots for F4/80highGr-1low peritoneal macrophages after magnetic bead enrichment of D5 post-injected peritoneal exudates. ! Figure S4. Itgb2-/- dendritic cells are hypersensitive to TLR stimulation. (A) and (B) Bone marrow-derived dendritic cells were isolated by

magnetic bead separation for MHC II+ cells after GM-CSF culture. DCs were stimulated with TLR agonists overnight and cytokine concentrations in the supernatant were determined by ELISA. The data are representative of 3 experiments and shown as mean +/- SD of independently stimulated triplicate wells. * p < 0.05. Figure S5. CD11a, CD11b, and Cbl-b deficiency selleck screening library does not induce macrophage TLR hypersensitivity or disturb MyD88 degradation. (A) Representative data of the results shown in Fig. 4A. WT, Itgal-/- (CD11a KO), Itgam-/- (CD11b KO) and Itgb2-/- macrophages were stimulated with 1 ng/mL LPS, 100 nM CpG DNA or 100 μg/mL zymosan particles for 24 hours and supernatant IL-12 p40 concentrations were determined by ELISA. Data are shown as mean +/- SD of independently LY2109761 stimulated triplicate

wells from one experiment. (B) Representative data of the results shown in Fig. 4C. Macrophages were stimulated as in (A) and cytokine concentrations were determined by ELISA. The results are displayed as mean +/- SD of independently stimulated wells from one experiment. (C) Macrophages were stimulated with 10 ng/mL LPS and cytoplasmic lysates were assessed for MyD88 by Western blot, with β actin used as a loading control. Results are representative of 2 independent experiments. ! Figure S6. β2 integrin deficiency enhances NF-κB pathway activation downstream of TLR activation. (A) and (C) Western blot analysis for macrophages stimulated with 1 ng/mL LPS for phospho-

IκBα, with β actin used as a loading control. In (A) and (C), macrophages were pre-treated with 10 μM MG-132 for 30 min. prior to LPS treatment. (B) and (D) Relative densitometry ratios (phospho-IκBα/β actin) for the data represented in (A) and (C) respectively. The results in (B) and (D) are set at Liothyronine Sodium WT time 0 set to 1 and shown as mean +/- SD of 2 separate experiments. (E) Macrophages were stimulated with 20 ng/mL TNF and expression of NF-kB-dependent genes was determined by qPCR, with results normalized to GAPDH expression and set relative to WT at time 4 hours. The results are shown as mean +/- SD of 2 independent experiments. ! “
“Natural killer (NK) cells form a region of tight contact called the NK immunological synapse (NKIS) with their target cells. This is a dynamic region serving as a platform for targeted signaling and exocytotic events. We previously identified IQGAP1 as a cytoskeletal component of the NK-like cell line YTS. The present study was undertaken to determine the role of IQGAP1 in the function of NK cells.