Fig. 1. Generation of model cell lines designed to study protein unfolding aspects during unconventional secretion of FGF-2. (A) cDNA constructs used in this study. CHOMCAT-TAM2 cells (Engling et al., 2002) were transduced with retroviral particles carrying the cDNA constructs shown. Clonal cell lines were obtained by FACS sorting. For details, see Materials and Methods. (B) Western blot analysis of model cell lines grown in the absence or presence of doxicycline. Cell lysates were separated on 10% SDS gels followed by protein transfer to a PVDF membrane. The constructs depicted in A were detected using affinity-purified anti-GFP antibodies (Engling et al., 2002). In the case of FGF-2-GFP, the material derived from about 10,000 cells was loaded; in the case of FGF-2-GFP-DHFR and MTS-GFP-DHFR, the material derived from about 20,000 cells was loaded.