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. 2011 Mar 14;6(3):e17922. doi: 10.1371/journal.pone.0017922

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Alfaro et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Transwell chemotaxis assays were set up with PKH2-labeled neutrophils. Neutrophils migrated to recombinant IL-8 added to the lower chamber. However, if neutrophils are seeded in the upper chamber together with DC (1∶1) migration is totally impaired. Of note there is IL-8 by the DC as shown in figure S6A. This experiment is representative of two independently performed in triplicate wells with migration lasting for 120 min. (B) Percentage of migration of PKH2-labeled neutrophils that were seeded into the upper chamber of transwell migration assays towards recombinant IL-8 or DC placed in the lower chamber. When indicated, IL-8-neutralizing mAb (20 µg/mL) was added to the lower chamber along with the DC. Migration was analyzed at 120 minutes. Similar experiments analyzed as early as 60 minutes rendered similar results (data not shown). Results are representative of two separate triplicate experiments independently performed. Asterisks indicate statistical significance p<0.01 in student's t tests.