Figure 1. CHO-cells express GFP-CD14 or PfRH5-CD14 inside cells and on their surface after transfection. (A) Representative cells expressing different recombinant proteins with GPI anchor in the endoplasmic reticulum/Golgi (ER) and partially on the cell surface. CHO cells transfected with pcDNA3-GFP-CD14-GPI were stained with ethidium bromide (EB, stains DNA) and ER-Tracker. For localization, the GFP fluorescence was observed at the same excitation for all markers, showing areas of colocalization between GFP and ER-tracker. DIC indicates the clear field image. (B) The same CHO GFP-CD14-expressing cells were stained with DAPI for visualizing nuclei, and GFP fluorescence is visible throughout the cell and on the cell surface. (C) Cells transfected with PfRH5-CD14 with 6xHis/Strep tags were stained with DAPI, whereas the presence of PfRH5-CD14 around the cells was highlighted by incubation with ant-6xHis as primary antibody and a secondary antibody anti-mouse IgG conjugated to Alexa 594. Wild type CHO cells showed no significant fluorescence when exposed to anti-6xHis and the anti-mouse Alexa 594 (data not shown).