Figure 4

Colocalization of APH0215-GFP with trans-Golgi network. (A) HeLa cells transfected with plasmid expressing APH0215-GFP, or GFP (green color), were subjected to immunofluorescence labeling with mouse monoclonal antibody against TGN38, a trans-Golgi network protein (red color). Arrowheads indicate the colocalization between APH0215-GFP and TGN38. The boxed area are magnified on the right. Scale bars: 5 μm. (B) The expression of APH0215-GFP and GFP in transfected cells. HeLa cells transfected with plasmid expressing APH0215-GFP (pAPH0215-GFP), or GFP (pEGFP-N1), were subjected to Western blot analysis using mouse monoclonal antibodies against GFP and β-tubulin (tubulin). Asterisks highlight the expected bands. Arrow indicates the degraded product of APH0215-GFP. β-tubulin is used as internal control for sample loading. (C) The expression level of TGN38 in transfected cells. HeLa cells transfected with plasmid expressing APH0215-GFP (pAPH0215-GFP), or GFP (pEGFP-N1), were subjected to Western blot analysis using mouse monoclonal antibodies against TGN38 and β-tubulin (tubulin). β-tubulin is used as internal control for sample loading. The full-length blots for all cropped ones are included in Supplemental Fig. S3 respectively.