TABLE 1.

Effects of pharmacological inhibitors on PMN cell surface protein expression

Damnacanthal treatment induces an up-regulation of CD11b/CD18, CD47, and SIRPα on the PMN surface. In these experiments, freshly isolated PMN (5 × 10⁶ cells) were incubated with damnacanthal or other inhibitors in their working concentrations for 15 min at 25 °C. After blocking with 5% normal goat serum, PMN cell surface CD11b, CD47, and SIRPα were labeled with LM2/1, PF3.1 (10 μg/ml each), and anti-SIRPα.ex (1:2000 dilution), respectively, for 1 h (4 °C). After washing and fixation with 3% paraformaldehyde (5 min, 20 °C), cells were incubated with Alexa Fluor-conjugated secondary antibodies followed by washing and analysis by flow cytometry. PMN labeled with normal mouse IgG served as base line. The table shows the mean fluorescence intensity ± S.E. (S.D. divided by the square root of the sample size).

Inhibitors	PMN surface fluorescence intensity (mean ± S.E.)
	Control IgG	CD11b/CD18	CD47	SIRPα
Vehicle control (DMSO)	3.2 ± 0.08	90.7 ± 0.11	35.2 ± 0.038	14.2 ± 0.09
Damnacanthal (10 μm)	2.5 ± 0.18	203.8 ± 0.22	112.8 ± 0.19	67.8 ± 0.16
Genistein (100 μg/ml)	4.3 ± 0.13	76.6 ± 0.28	30.0 ± 0.13	19.6 ± 0.14
Piceatannol (40 μm)	5.5 ± 0.26	94.0 ± 0.17	28.2 ± 0.14	13.5 ± 0.16
PP1 (18 μm)	2.7 ± 0.33	85.2 ± 0.43	29.3 ± 0.46	11.6 ± 0.31
LFM-A13 (140 μm)	4.4 ± 0.51	89.3 ± 0.44	28.5 ± 0.69	9.7 ± 0.42
Herbimycin A (2 μm)	5.2 ± 0.26	95.7 ± 0.25	30.3 ± 0.15	16.3 ± 0.45
Lavendustin A (10 μm)	5.5 ± 0.16	91.4 ± 0.32	32.2 ± 0.21	20.1 ± 0.25
Erbstatin analog (25 μm)	4.8 ± 0.17	81.6 ± 0.20	29.8 ± 0.19	18.3 ± 0.23
SB203580 (1 μm)	6.1 ± 0.30	83.8 ± 0.40	28.8 ± 0.15	15.2 ± 0.23
PD98059 (80 nm)	3.7 ± 0.16	89.9 ± 0.24	26.7 ± 0.18	12.5 ± 0.25
LY294002 (100 nm)	3.0 ± 0.14	79.6 ± 0.13	31.9 ± 0.30	19.1 ± 0.15
AG126 (150 μm)	4.3 ± 0.17	97.5 ± 0.19	30.7 ± 0.14	14.6 ± 0.24