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. 2006 Apr 6;35(3):378–386. doi: 10.1165/rcmb.2006-0027OC

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Figure 6. — Effect of IL-5 on αMβ2 conformation, adhesion, and podosome formation of purified blood EOS. (A) Purified blood EOS from normal or atopic subjects were incubated without (thin line) or with (bold line) 50 ng/ml IL-5 for 20 min at 37°C and then placed on ice before addition of CBRM1/5. Similar increases in CBRM1/5 reactivity upon IL-5 treatment of blood EOS were found in four separate donors, P = 0.0417, two-tailed t test. Unchallenged blood EOS from normal or atopic subjects were incubated with 50 pg/ml IL-5 for 5 min before addition of cells to wells coated with (B) 10 μg/ml diverse ligands or (C) 1% BSA in the presence of inhibitory mAbs. Results are the mean and SD of assays performed in triplicate for five separate donors (15 wells). *P < 0.001, comparing adhesion on ligands to the FBS blocker or adhesion in the presence of inhibitory mAbs to adhesion with respective isotype control mAbs, one-way ANOVA with Dunnett's post test. (D) Blood EOS treated with 100 ng/ml IL-5 and adherent to VCAM-1 were assayed for podosome formation by immunofluorescent staining of actin with rhodamine-phalloidin. Scale bar = 10 μm.

Figure 6. — Effect of IL-5 on αMβ2 conformation, adhesion, and podosome formation of purified blood EOS. (A) Purified blood EOS from normal or atopic subjects were incubated without (thin line) or with (bold line) 50 ng/ml IL-5 for 20 min at 37°C and then placed on ice before addition of CBRM1/5. Similar increases in CBRM1/5 reactivity upon IL-5 treatment of blood EOS were found in four separate donors, P = 0.0417, two-tailed t test. Unchallenged blood EOS from normal or atopic subjects were incubated with 50 pg/ml IL-5 for 5 min before addition of cells to wells coated with (B) 10 μg/ml diverse ligands or (C) 1% BSA in the presence of inhibitory mAbs. Results are the mean and SD of assays performed in triplicate for five separate donors (15 wells). *P < 0.001, comparing adhesion on ligands to the FBS blocker or adhesion in the presence of inhibitory mAbs to adhesion with respective isotype control mAbs, one-way ANOVA with Dunnett's post test. (D) Blood EOS treated with 100 ng/ml IL-5 and adherent to VCAM-1 were assayed for podosome formation by immunofluorescent staining of actin with rhodamine-phalloidin. Scale bar = 10 μm.