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. Author manuscript; available in PMC: 2011 Nov 1.
Published in final edited form as: Nat Immunol. 2008 Dec 7;10(1):101–108. doi: 10.1038/ni.1675

Figure 2.

Figure 2

DARC immunoreactivity in MDCK-DARC monolayers. (a–c) Confocal laser-scanning microscopy of the subcellular distribution of DARC immunoreactivity in cross-sections of unstimulated (a) and CCL2-stimulated (b,c) MDCK-DARC monolayers grown on Transwell inserts; in b,c, 35 nM CCL2 was added below the monolayers, followed by incubation for 30 min (b) or 2 h (c). DARC immunoreactivity was detected by indirect immunofluorescence with Alexa Fluor 488 (quantification of DARC distribution, Supplementary Table 1). Scale bars, 10 μm. (d) Immunoelectron microscopy of DARC immunoreactivity (electron-dense dots of colloidal gold) on apical microvillus extensions of MDCK-DARC cells 2 h after the application of CCL2 to the basolateral side. Arrows indicate apical microvilli. Scale bar, 500 nm. (e) Immunoelectron microscopy of the colocalization of DARC (large black dots; 15-nm colloidal gold) with CCL2 (small black dots; 5-nm colloidal gold) on an apical microvillus extension (arrow). Scale bar, 200 nm. Images in d,e were acquired with a transmission electron microscope. Results are representative of at least three independent experiments.