Fig. 3

Adhesion of Mac-1-expressing cells to Dyn A and Dyn B. a, Mac-1-expressing and wild-type HEK293 cells were labeled with calcein and 5x10⁴ cells were added to the wells coated with different concentrations of the Dyn A. After 30 min at 37 °C, nonadherent cells were removed and fluorescence was measured. The data shown are means of triplicate determinations, and error bars represent S.E. The figure is representative of 8 experiments. b , adhesion of HEK293 cells expressing Mac-1, LFA and the I-less form of Mac-1 to Dyn A and Dyn B immobilized at 10 μM was determined as described in A. c, calcein-labeled Mac-1-HEK293 cells were preincubated without or with 10 μg/ml of anti-α_M mAb 44a or 1 μg/ml NIF for 15 minutes. For testing the role of glucosaminoglycans in cell adhesion, dynorphin A-coated wells were preincubated with 100 μl of 10 μg/ml of chondroitin sulfate A, chondroitin sulfate B and heparin. Aliquots (5x10⁴cells) were added to the wells coated with 2 μM dynorphin A and adhesion was performed as described in A. Adhesion in the absence of Mac-1 inhibitors and glucosaminoglycans was assigned a value of 100 %. The data shown are the means ± S.E. from four separate experiments each with triplicate measurements. d , microtiter plate wells were coated with different concentrations of dynorphin B and adhesion of Mac-1-expressing and wild-type HEK293 cells was determined as described in A. The data shown are means of triplicate determinations, and error bars represent S.E. The figure is representative of 5 experiments. e. calcein-labeled Mac-1-HEK293 cells were preincubated without or with 10 μg/ml of mAb 44a and heparin, added to the wells coated with 20 μM of dynorphin B and adhesion was performed as described in A. Adhesion in the absence of inhibitors was assigned a value of 100 %. The data shown are the means ± S.E from three separate experiments each with triplicate measurements. **p < 0.01; ns, no significant difference