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. 2015 Mar 30;10(3):e0122253. doi: 10.1371/journal.pone.0122253

Fig 6. Improved utility of the inducible, bidirectional system by transposon delivery of the tetracycline repressor protein.

Fig 6

(A) Sleeping Beauty (SB) transposons encoding for inducible expression of influenza A virus hemagglutinin (HA) gene (HA-IE-N) or bicistronic expression of the tetracycline-repressor (TetR) and puromycin resistance gene (Puro) were cotransfected with SB transposase (PGK-SB11) into HeLa cells to create cell lines with regulated levels of HA. (B) Western blot of total cell lysates prepared from two cell lines cultured in the absence of doxycycline (repressed, R), presence of 4 μM doxycycline (de-repressed, DR), or when doxycycline treated cells were transduced with adenovirus vector particles (m.o.i. = 3) encoding for expression of VP16 (induced, IN). Membranes were reacted with antibodies to HA or GAPDH, which served as a loading control. Molecular weights (kDal) are indicated. PGK, human phosphoglycerate kinase promoter; Cags, chimeric CMV enhancer:chicken beta-actin promoter; ires, internal ribosome entry site; pA, BGH polyadenylation signal; A; SV40 polyadenylation signal.