|
|
Blood, Vol. 106, Issue 13, 4241-4248, December 15, 2005
Costimulation of mast cells by 4-1BB, a member of the tumor necrosis factor receptor superfamily, with the high-affinity IgE receptor
Blood Nishimoto et al.
106: 4241
Supplementary Figures for: Nishimoto et al
Files in this Data Supplement:
- Figure S1. Induction of 4-1BB expression on mast cells by IL-3, SCF, and LPS (JPG, 45.9 KB)
-
Naïve (non-IgE sensitized) B6 BMMC were cultured without IL-3 for 6 h, and then incubated with 100 ng/ml of IL-3 or 100 ng/ml of SCF for 6 h (A), or 1 µg/ml of LPS for 12 h (B) before flow cytometric analysis for surface expression of 4-1BB. Results shown are representative of two experiments.
- Figure S2 (JPG, 38.1 KB)
-
Levels of cytokine mRNAs induced by IgE/Ag stimulation are reduced in 4-1BB-/- mast cells. Wt and 4-1BBL-/- BMMC were left unstimulated (-) or incubated overnight with anti-DNP IgE (206). The IgE-sensitized cells were washed and incubated with 10 ng/ml DNP23-HSA for the indicated periods. RNAs were isolated and subjected to semi-quantitative RT-PCR for the expression of IL-6, IL-13, and GAPDH mRNAs. Results shown are representative of two experiments.
- Figure S3 Cell surface phenotypic, morphological, and growth properties of 4-1BBL-/- mast cells (JPG, 51.7 KB)
-
(A) Wt and 4-1BBL-/- BMMC were generated from bone marrow cells in IL-3-containing culture medium. Cohorts of 3-5 mice in each group were used to generate BMMC. Results shown in this figure are representative of two independent experiments. Surface expression of Fc RI and c-Kit was analyzed by flow cytometry. (B) Toluidine blue-stained wt and 4-1BBL-/- mast cells. (C) Growth curves of bone marrow cells derived from wt and 4-1BBL-/- mice in IL-3-containing medium. (D) Wt and 4-1BBL-/- mast cells that had been deprived of growth factors for 8 h were incubated with the indicated concentrations of IL-3 and SCF for 12 h. DNA synthesis was measured by [3H]thymidine incorporation during the last 6 h of cultures. (E) Wt and 4-1BBL-/- mast cells were incubated without IL-3 or other growth factors for the indicated periods. Live cells were quantified by flow cytometry of annexin V- and propidium iodide-stained cells. Annexin V-negative and propidium iodide-negative cells are plotted as function of incubation time.
|
|
