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MATERIALS & METHODS

MICROSCOPIC TECHNIQUES SPECIES MORPHOLOGY CELL LINE
621 Localization and functional interrelationships among cytosolic Group IV,
secreted Group V, and Ca2+-independent Group VI phospholipase A2s in
P388D1 macrophages using GFP/RFP constructs
Yasuhito Shirai, Jesus Balsinde, Edward A. Dennis phospholipase A2, arachidonic acid, macrophage, platelet-activating factor, lipopolysaccharide, prostaglandin

An appropriate number of P388D1 cells transiently or stably expressing the PLA2 fusion proteins were placed on a glass-bottomed dish (MetTek, Ashland, MA) and cultured overnight.

epifluorescence microscopy, confocal microscopy P388D1
  P388D1 cells exposed to bacterial lipopolysaccharide (LPS) mobilize arachidonic acid (AA) for prostaglandin synthesis in two temporally distinct pathways. The ‘‘immediate pathway’’ is triggered within minutes by receptor agonists such as platelet-activating factor (PAF) but only if the cells have previously been primed with LPS for 1 h. The ‘‘delayed pathway’’ occurs in response to LPS alone over the course of several hours. We have now investigated  

 

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