Determination of fate of CCR2 downregulation on monocytes from Ficoll.

CCR2 expression was assessed after Ficoll isolation of untagged and tagged-CCR2 monocytes. Internalization of the complex was examined from one single layer of Z stacks by confocal microscopy. The results of a representative experiment are shown: a) CCR2 expression on monocytes from WB tagged with CCR2 prior to gradient separation; b) CCR2 expression on untagged monocytes after Ficoll isolation and the subsequent staining of CCR2; c) CCR2 expression in CCR2 tagged monocytes after Ficoll isolation, red signal within monocytes represents the CCR2-complex internalization (no aditional anti CCR2-PE was added to the cells after isolation with Ficoll); (isotype control □ , chemokine receptor ▪).