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. Author manuscript; available in PMC: 2011 Apr 11.
Published in final edited form as: J Immunol. 2010 Mar 31;184(9):5232–5241. doi: 10.4049/jimmunol.0902501

FIGURE 4.

FIGURE 4

sPLA2-induced production of VEGF-A is independent from enzymatic activity and involves a receptor-mediated activation of HLMs. A, Effect of H48Q mutants of sPLA2s on VEGF-A release. HLMs were incubated (37°C, 24 h) with RPMI 1640 alone (ctr), with the wt hGIIA and hGX, or the mutants H48Q of hGIIA and hGX. VEGF-A release was determined by ELISA. Data are mean ± SEM of three experiments. *p < 0.05 versus control. B, Effect of Me-Indoxam and RO092906A on hGX-induced VEGF-A release. hGX (3 μg/ml) was preincubated (37°C, 15 min) with RPMI 1640 alone, or with the indicated concentrations of Me-Indoxam (●) or RO092906A (○). HLMs were then incubated (37°C, 24 h) with hGX (3 μg/ml) alone or with the various combinations of hGX with Me-Indoxam (●) or RO092906A (○). VEGF-A release was determined by ELISA. Data are expressed as percent inhibition of the maximum response induced by hGX alone calculated as (R – Rb)/(Rmax – Rb) × 100, where R is the release in samples treated with the combination hGX plus inhibitor, Rb is the release in unstimulated samples, and Rmax is the release in samples stimulated with hGX alone. Data are the mean ± SEM of three experiments. VEGF-A release induced by hGX alone was 661 ± 83 pg/mg protein. The unbroken and broken lines represent the best fit for inhibition of Me-Indoxam and RO092906A, respectively. C, Effect of MAPK inhibitors on sPLA2-induced VEGF-A release. The cells were preincubated (37°C, 1 h) with SB203580 (30 μM), PD98059 (50 μM), or both (30 μM SB203580 and 50 μM PD988059) and then stimulated (37°C, 24 h) with hGX (3 μg/ml). VEGF-A release was determined by ELISA. Data are the mean ± SEM of three experiments. *p < 0.05 versus control; §p < 0.05 versus hGX alone.