FIGURE 1.

Expression of various intracellular PLA₂ in mast cells. A, RNAs obtained from BMMCs maintained in IL-3 and those cocultured for the indicated periods with Swiss 3T3 cells (CTMC-like cells) were subjected to RT-PCR for various PLA₂s (28 cycles). RT-PCR for mMCP-4, a CTMC-specific protease, was performed as a positive control for the differentiation of immature BMMCs into mature CTMCs. Equal loading of each sample was verified by RT-PCR of Gapdh, a housekeeping gene. RT-PCR without RNA (H₂O) was done as a negative control. B, quantitative RT-PCR of intracellular PLA₂s in BMMCs. C, lysates of replicate cells were subjected to SDS-PAGE (8% gel) followed by immunoblotting with cPLA₂α and iPLA₂β. Arrows indicate the positions of major bands for individual PLA₂s. D, effects of (S)- or (R)-BEL on β-HEX release from IgE/Ag-activated BMMCs (mean ± S.E., n = 5; *, p < 0.05 and **, p < 0.01).