Fig. 5. The effect of BFA on the trafficking of mutant Cx32I28L. COS-7 cells were transfected with cDNA encoding Cx32I28L and 48 hours post-transfection cells were fixed and stained with an antibody against the carboxyl tail of Cx32 (Gap34R). (A) Cx32I28L under normal conditions. Similar images were obtained for wtCx32 and mutant Cx32V206I. (B) Mutant Cx32I28L 6 hours post-treatment with BFA. Arrow shows retention of gap junctions at the plasma membrane. (C) Cx32V206I 6 hours post treatment with BFA. Note that Cx32 staining was intracellular. (D) wtCx26 6 hours following nocodazole treatment showing that Cx26 was intracellular. (E) wt Cx32 6 hours following nocodazole treatment. Arrow shows gap junctions. (F) Mutant Cx32I28L 6 hours following nocdoazole treatment showing limited plasma membrane but much increased intracellular staining. D and F show a reduction in plasma membrane staining. Bar, 10 µm, Arrows indicate targeting to the gap junction. Parallel staining of the cells with p58 or ß-tubulin was used to confirm that disassembly of the Golgi following BFA treatment and shattering of the microtubules following nocodazole treatment had occurred in a similar manner to that shown in Fig. 3 (not shown).
