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. Author manuscript; available in PMC: 2015 Jun 2.
Published in final edited form as: Oncogene. 2009 Apr 20;28(21):2163–2172. doi: 10.1038/onc.2009.82

Figure 4.

Figure 4

Effects of inducible dominant-negative TCF-4 (DNTCF-4) on growth of non-small-cell lung carcinoma (NSCLC) autocrine cells. (a) Immunoblot analysis of DNTCFs expression. H23 and H1819 cells were infected with lentiviruses expressing DNTCF-4 (DN), DN-mOrange (DN-mO) and vector (VEC) under the control of a tetracycline-inducible promoter and selected with puromycin in the presence of dox. After washing, cells were divided into separate cell culture dishes in the presence or absence of dox and analysed by immunoblot 3 days after induction. Expression of DNTCF-4 proteins was detected using an antibody to TCF-4. Lower molecular weight immunoreactive DNTCF species were also observed. Molecular weights in kilodaltons are indicated (kDa). (b) Effect of DNTCFs on axin2 mRNA expression. RNA was extracted from H23 and H1819 cells infected as in (a) and maintained in the presence or absence of dox for 3 days. (c) Effect of DNTCFs on cell-cycle profile. Propidium iodide (PI) analysis of H23 and H1819 cells infected as in (a) and maintained in the presence or absence of dox at 3 days after induction. Numbers indicate the percentage of cells in G1 or S phase for each cell line analysed. Results are representative of at least two independent experiments. (d) Effects on proliferation of H23 cells expressing inducible DN-mO in the presence or absence of dox, observed at 3 days after induction. BF, bright field; FL, fluorescence. (e) Effects on growth of H23 and H1819 at 2–3 weeks following expression of VEC, DN and DN-mO. A total of 2 × 104 cells were plated into 60mm plates in the presence or absence of dox. Cultures were visualized by crystal violet staining. (f) Effects of DNTCFs on expression of c-Myc, cyclin D1 and p21. Protein lysates from H23 and H1819 cells, infected with VEC, DN and DN-mO, and grown in the presence or absence of dox for 3 days, were analysed by immunoblot. Dox, doxycycline.