Fig. 4.

IFN reversibly displaces Cx43 from lipid rafts in IEC-6 cells and decreases the phosphorylation of Cx43 at lipid rafts; these events are reversed by PMA. A: IEC-6 cells were either untreated (i–iii), treated with IFN (iv–vi) or treated with IFN in the presence of PMA (vii–ix) as described in methods. Cells were stained with antibodies against Cx43 (red staining) and Cav-1 (green staining) along with the nuclear marker Draq-5 (blue staining) and were examined by immunoconfocal microscopy. Arrows indicate sites of colocalization of Cx43 and Cav-1 on the merged images. Representative of 5 separate experiments. B: confocal micrographs of IEC-6 cells that were either untreated (i) or were treated with IFN in the absence (ii) or presence (iii) of PMA, treated with cholera toxin and immunostained with antibodies against phosphorylated Cx43 as described in methods. The colocalization of phosphorylated Cx43 with lipid rafts is revealed by arrows. Representative of 3 separate experiments.