Figure 4. Localization by immunofluorescence of the Ins(1,4,5)P₃R₂ in rat hepatocytes grown for 1 and 4 h in primary culture.

The culture of rat hepatocytes, permeabilization and cell fixation, staining with anti-Ins(1,4,5)P₃ antibody, and confocal microscopy were performed as described in the Materials and methods section. Cells were stained with mouse monoclonal anti-Ins(1,4,5)P₃R₂ antibody KM1083 and anti-mouse antibody conjugated to Cy3. The results shown are those obtained for one of three experiments employing three separate rat hepatocyte preparations which each gave similar results. The scale bar represents 10 μm. Panels a–f represent different cells showing the bright-field images (a, c and e) and immunofluorescence (equatorial sections) (b, d and f) for cells cultured at 1 h. Panels h–m represent different cells showing the bright-field images (h, j and l) and immunofluorescence (equatorial sections) (i, k and m) for cells cultured at 4 h. Panels g and n are controls in which the primary antibody has been omitted.