Fig. 5. Expression of fusion proteins between teneurin-2 and the recombinant transcription factor BDAD. Western blot analysis (A) and immunofluorescence staining (B) of BDAD and three different BDAD-teneurin-2 fusion constructs in HT1080 cells using anti-Gal4 antibodies. (A) Transient transfection with constructs pBD-NFB expressing BDAD, BDAD-I, BDAD-ITE and BDAD-ITEY in the presence (+) or absence (-) of ALLN, as indicated above the lanes. (B) Immunofluorescence analysis of permeabilized cells transfected with BDAD-I and of non-permeabilized cells with either BDAD-ITE or BDAD-ITEY using anti-Gal4 to detect BDAD-I and anti-teneurin-2 for BDAD-ITE and BDAD-ITEY. Staining of BDAD-ITE- or BDAD-ITEY-transfected cells with anti-Gal4 after permeabilization resulted in addition to the cell surface staining in ER and Golgi staining, but no nuclear staining could be detected (data not shown). BDAD-I did not accumulate in PML bodies but was diffusely distributed throughout the nucleus and was strongly protected from degradation in the presence of the proteasome inhibitor ALLN. Bar, 20 µm.