Regulation of Hemichannels and Gap Junction Channels by Cytokines in Antigen-Presenting Cells

Figure 6

B cells present pannexin1 at the cell surface. Confocal images (Olympus, FluoView FV1000) of immunofluorescence analysis of freshly isolated B cells fixed in ethanol (70%). B cells were isolated from peripheral lymph nodes by positive selection from balb/c mice. Top left: B cells were identified with IgG (conjugated to FITC, green); the inset shows the bright field. Top right: pannexin1 (Panx1) immunoreactivity (red, primary antibody: rabbit anti-Panx1 antibody and secondary antibody goat anti-rabbit IgG conjugated to Cy3) is shown. Bar: 20 μm. Middle left: using ImageJ colocalization finder, it can be seen that Panx1 colocalizes with IgG (white) at the cell surface in some B cells (white arrows). B cells with low or no colocalization are indicated (green arrows). Middle right: zoom and merge of IgG and Panx1 labeling in a B cell denoted by a dotted square at middle left panel. The white line denotes the region used for the line scan. Bar: 10 μm. Bottom: ImageJ line scan analysis shows the fluorescence intensity of each channel through the white line in the middle of each cell. The peak coincidence (denoted by dotted squares) is an index of colocalization between the different fluorophores.