In polarized CFPAC-1 cells, osmium impregnation revealed a large number of Golgi stacks dispersed throughout the cytoplasm (Figure 2a 1). Each Golgi stack contained chromophilic (Figure 2a 2, arrow) and chromophobic (Figure 2a 2, arrowhead) regions corresponding to the cis and trans faces, respectively. They often selleck screening library appeared in groups of two or three (Figure 2a 1, arrows). Electron microscopic examination confirmed this dispersed stack distribution. Figure 2b shows a polarized CFPAC-1 cell containing many small Golgi stacks disconnected from one another and scattered throughout the cytoplasm (Figure 2b, arrows). These stacks were unusual in that they were formed by dilated cisternae, often highly vesiculated (Figure 2c).

In polarized CFPAC-PLJ-CFTR6 cells, Golgi stacks revealed by osmium impregnation were mostly grouped in the supranuclear cytoplasm (Figure 2d, arrows). Ultrastructural analysis showed a well-organized Golgi complex composed of large stacks with flattened cisternae (Figure 2e). Confocal microscopic examination of the immunocytochemical reactions using anti-58K protein antibodies demonstrated the presence of numerous Golgi stacks dispersed throughout the cytoplasm in polarized CFPAC-1 cells: (a) in focal planes passing through the bases of cells, few Golgi stacks were present (Figure 3a 1); (b) in medial planes, a large number of stacks either isolated, or in small clusters (Figures 3a 2 and and3a3a 3, arrows) appeared throughout the cytoplasm; and (c) in supranuclear planes, there were very few or none at all (Figure 3a 4).

In polarized CFPAC-PLJ-CFTR6 cells, the distribution of Golgi stacks was different: they were absent in the basal cytoplasm (Figure 3b 1), few in number in the medial cytoplasmic regions (Figure 3b 2), and numerous and grouped in clusters in the supranuclear regions (Figures 3b 3 and and3b3b 4, arrows). We determined the number of cells presenting dispersed, partially dispersed, or clustered Golgi stacks using immunofluorescence images of different planes taken along the entire height of cells. For each cell line, we carried out counts of more than 200 cells per preparation (n=5). Statistical analyses of these data were performed using the non-parametric Mann-Whitney test. In the CFPAC-1 line, 90.1 �� 1.5% of the cells exhibited a dispersed and 4.1 �� 0.6% a clustered Golgi complex vs 12.4 �� 2.5% and 73.5 �� 4.

1% in the CFPAC-PLJ-CFTR6 cell line (p<0.0001). A partially dispersed Golgi complex was observed in 5.7 �� 1.1% of CFPAC-1 cells and 14 �� 3% of CFPAC-PLJ-CFTR6 cells. Figure 2. Characteristics of the Golgi complex in CFPAC-1 (a�Cc) and CFPAC-PLJ-CFTR6 (d,e) cells. (a1) Dispersal of the Golgi complex Entinostat revealed by osmium impregnation. The arrows indicate the presence of associations between different Golgi stacks. Bar = … Figure 3.