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Fig. 5. Two distinct signaling pathways in LPC-induced azruophil granule translocation. (A) Anti-G2A Ab blocks LPC-induced CD63 surface expression. Neutrophils were pretreated with anti-G2A Ab (1 μg/ml) for 1 h and further stimulated with LPC (3 μM) for 15 min. Then neutrophils were fixed and stained with FITC-conjugated anti-CD63 Ab for 1 h on ice. (B, C) SB203580 blocks LPC-induced CD63 surface expression, but not F-actin polymerization. Neutrophils were pretreated with SB203580 (10 μM) for 30 min, and further stimulated with LPC (3 μM) for 15 min. After fixation, cells were stained with FITC-conjugated anti-CD63 Ab (B) or FITC-conjugated phalloidin (C) for 1 h. (D) Y27632 does not affect LPC-induced [Ca2+]i increase. Neutrophils were loaded with Fluo-3 AM (4 μM) in HEPES for 1 h and, then pretreated with Y27632 (10 μM) for 30 min before LPC stimulation. [Ca2+]i changes in Fluo-3 AM-loaded neutrophils were expressed as the relative fluorescence intensity over the resting fluorescence value (F/Fo). An average ± SEM of more than three experiments is shown. LPC, lysophosphatidylcholine; HEPES, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; MFI, mean fluorescence intensity. *p < 0.05, **p < 0.01, and ***p < 0.001.
Korean J Physiol Pharmacol 2022;26:175-182 https://doi.org/10.4196/kjpp.2022.26.3.175
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