Figure 3
Figure 3. GC-treated cells show weaker adhesion as well as increased migratory and chemotactic activity. (A) Monocytes were treated with 10 nM fluticasone (GC) or left untreated (CO) for 2 days in Teflon bags and subsequently allowed to adhere to multiwell plates for 2 hours. After removing nonadherent cells by washing, remaining adherent cells were detached with EDTA and counted in a cell counter. The complete detachment of adherent cells was confirmed microscopically. The bars represent mean and SEM of 3 independent experiments. The decrease in adherence was statistically significant (*P < .05). (B) Expression of formyl peptide receptor (FPR), which was found to be differentially expressed by microarray analysis, was confirmed by flow cytometry. Monocytes were treated as described in panel A. Specific profiles of FPR expression are shown by thick lines, and isotype controls appear as spotted lines. Numbers indicate the quotient of specific/isotype control MFI and percentage of positive cells, respectively. The experiment was done 3 times with similar results. The increase in MFI was calculated to be statistically significant (P < .05, Student t test). (C) Monocytes were treated as described in panel A and placed into the upper chamber of a transwell filter. The lower chamber contained monocyte medium without or with increasing concentrations of fMLP as chemotactic stimulus. After 4 hours the number of cells which had migrated into the lower compartment was counted. The bars represent mean and SEM of 3 independent experiments. The increase in chemotactic activity was statistically significant (*P < .05, **P < .01). (D) Monocytes were treated as described in panel A and placed into the upper chamber of a transwell filter. The lower chamber contained monocyte medium with the addition of 10 nM fMLP, 100 nM LTB4, or no attractants but 10 nM fMLP in the upper chamber. After 4 hours cells that had migrated into the lower compartment were counted, and numbers are presented as the percentage of cells which migrated in the absence of any chemotactic stimulus. The bars represent mean and SEM of 3 independent experiments. The increase in chemotactic activity was calculated to be statistically significant (**P < .01).

GC-treated cells show weaker adhesion as well as increased migratory and chemotactic activity. (A) Monocytes were treated with 10 nM fluticasone (GC) or left untreated (CO) for 2 days in Teflon bags and subsequently allowed to adhere to multiwell plates for 2 hours. After removing nonadherent cells by washing, remaining adherent cells were detached with EDTA and counted in a cell counter. The complete detachment of adherent cells was confirmed microscopically. The bars represent mean and SEM of 3 independent experiments. The decrease in adherence was statistically significant (*P < .05). (B) Expression of formyl peptide receptor (FPR), which was found to be differentially expressed by microarray analysis, was confirmed by flow cytometry. Monocytes were treated as described in panel A. Specific profiles of FPR expression are shown by thick lines, and isotype controls appear as spotted lines. Numbers indicate the quotient of specific/isotype control MFI and percentage of positive cells, respectively. The experiment was done 3 times with similar results. The increase in MFI was calculated to be statistically significant (P < .05, Student t test). (C) Monocytes were treated as described in panel A and placed into the upper chamber of a transwell filter. The lower chamber contained monocyte medium without or with increasing concentrations of fMLP as chemotactic stimulus. After 4 hours the number of cells which had migrated into the lower compartment was counted. The bars represent mean and SEM of 3 independent experiments. The increase in chemotactic activity was statistically significant (*P < .05, **P < .01). (D) Monocytes were treated as described in panel A and placed into the upper chamber of a transwell filter. The lower chamber contained monocyte medium with the addition of 10 nM fMLP, 100 nM LTB4, or no attractants but 10 nM fMLP in the upper chamber. After 4 hours cells that had migrated into the lower compartment were counted, and numbers are presented as the percentage of cells which migrated in the absence of any chemotactic stimulus. The bars represent mean and SEM of 3 independent experiments. The increase in chemotactic activity was calculated to be statistically significant (**P < .01).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal