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. 2013 Sep 20;8(9):e75084. doi: 10.1371/journal.pone.0075084

Figure 5. The B7 cytoplasmic domain can accelerate the intracellular transport of a chimeric AFP protein.

Figure 5

a) AFP-PDGFR-B7 has the B7 cytoplasmic domain appended to AFP-PDGFR, in which AFP replaces the scFv in scFv-PDGFR (Figure 1c). b) 3T3 fibroblasts that were transiently-transfected with plasmids coding for AFP-PDGFR or AFP-PDGFR-B7 were immunofluorescence stained for the HA epitope tag on AFP. c) 3T3 cells expressing AFP-PDGFR or AFP-PDGFR-B7 were pulse-chased for the indicated times before surface (S, lower panel) and intracellular (I, upper panel) AFP chimeric proteins were immunoprecipitated, separated by SDS PAGE and visualized by autoradiography. Vector-transfected control cells are indicated as pHook-1. d) The percentage of intracellular Golgi-glycosylated AFP chimera in relation to total intracellular AFP chimeric protein in 35S-methionine labeled cells at the indicated chase times are shown.