Figure 2.

Overexpression of Different Arf6 Forms Do Not Affect ACTH Vesicle Transport into the Processes of AtT20 Cells

A, Immunocytochemistry showing the intracellular distribution of ACTH in AtT20 cells expressing RFP (top panels) or RFP-CPE_C10 (bottom panels). Scale bar, 5 μm. B, Immunocytochemistry showing the intracellular distribution of ACTH in AtT20 cells expressing HA-tagged wt/DN/CA-Arf6. ACTH and HA-tagged Arf6s were detected as described in Materials and Methods. Scale bar, 5 μm. C, Bar graph showing the quantification of the number of cells with accumulation of punctate ACTH immunostaining in the processes in untransfected cells (untransf.) and cells transfected with wt-Arf6, DN-Arf6, CA-Arf6, GFP, or GFP-CPE_C10. A total of 100 cells expressing each of the constructs was scored in three independent experiments. D, Bar graph showing the ratio of the mean immunostaining intensity of ACTH in the processes relative to the cell body in untransfected cells (untransf.) or cells overexpressing wt/DN/CA-Arf6, GFP/GFP-CPE_C10, or RFP/RFP-CPE_C10 (n = 40). The intensity of ACTH immunostaining in the processes and cell body of AtT20 cells in each condition was quantified using Metamorph software.