FIG. 6.

Primary-cell preparations, NCI-N87 cells, and AGS cells immunostained for α-dystroglycan and paxillin. (A to F) Cells were grown on APES-coated coverslips for 48 h prior to fixation with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. All cells were immunostained separately with monoclonal anti-α-dystroglycan and monoclonal antipaxillin antibodies for 18 h at 4°C. (G) α-Dystroglycan protein expression levels within primary-cell preparations, NCI-N87 cells, and AGS cells were analyzed by SDS-PAGE. (H) Paxillin protein expression levels within primary-cell preparations, NCI-N87 cells, and AGS cells were analyzed by SDS-PAGE. The housekeeping protein histone 2B was used to normalize for protein loading. The primary (A) and NCI-N87 (B) cells showed a strong immunostaining of the ER and Golgi complex (arrows), whereas the AGS cells (C) showed staining in intracellular vesicles (arrows). The levels of α-dystroglycan protein expression are highest in primary cells, followed (in order) by NCI-N87 and AGS cells (G). The primary cells (D) showed both cytosolic and focal contact staining, while in the NCI-N87 (E) and AGS (F) cells paxillin IR was predominantly at the focal contacts. The largest amount of paxillin protein was present in the primary cells, followed (in order) by AGS cells and NCI-N87 cells (H). Scale bar = 10 μm. Images are representative of three independent experiments.