Journal: PLoS ONE
Article Title: IL-33 Is Produced by Mast Cells and Regulates IgE-Dependent Inflammation
doi: 10.1371/journal.pone.0011944
Figure Lengend Snippet: 1×10 6 BMMC were primed with 1 µg/ml anti-DNP IgE (SPE-7 clone) overnight and stimulated by addition of 0.5 µg/ml DNP-HSA (black bars) or vehicle control (white bars). After the indicated times, gene expression of mouse IL-33 was determined by real-time RT-PCR (A). Either anti-DNP IgE or anti-OVA IgE was used in combination with either DNP-HSA or OVA, as indicated, and IL-33 gene expression determined (B). Intracellular staining for IL-33 was determined in permeabilized BMMC 24 hours after stimulation of 0.5 µg/ml DNP-HSA (C). Western blot analysis of BMMC lysate from stimulated or unstimulated cells 24 hours after stimulation (D). Recombinant, mouse IL-33 109–266 was used as a positive control. Data represents 3 independent experiments. Primary mast cells were obtained by peritoneal lavage, identified as CD117 + /FcεRI + and intracellular staining for IL-33 performed after permeabilization (E). Data represents 2 independent experiments.
Article Snippet: Rabbit polyclonal anti-IL-33 (Axxora) (1∶500) was used as a primary detection antibody and biotinylated goat anti-rabbit IgG (Zymed) was used as a secondary antibody.
Techniques: Expressing, Quantitative RT-PCR, Staining, Western Blot, Recombinant, Positive Control