Figure 2: Cell transformation and oncogenic effect of UVRAGFS mutant. | Nature Communications

Figure 2: Cell transformation and oncogenic effect of UVRAGFS mutant.

From: Truncating mutation in the autophagy gene UVRAG confers oncogenic properties and chemosensitivity in colorectal cancers

Figure 2

(a) NIH3T3 cells stably expressing empty vector, UVRAGWT, and UVRAGFS (104) were seeded and counted over time in triplicate. Values are the means±s.d. (n=4). Flag-tagged UVRAG expression is shown by western blot and β-tubulin serves as a loading control. *P<0.05; **P<0.01. (b) NIH3T3 cells described above were plated at low density (2,500 cells per 10-cm plate), grown for 14 days then fixed and stained with crystal violet. (c) Anchorage-independent growth induced by UVRAGFS. NIH3T3 cells in (a) were seeded in 0.3% top agar and incubated for 20 days. UVRAGFS-expressing cells formed larger and greater number of colonies in soft agar. Representative images of colonies are shown and the quantitative results of colony numbers were obtained from 10 randomly chosen HPF. Data represent the means±s.d. (n=4). **P<0.01; ****P<0.0001. Scale bar, 500 μm. (d) UVRAGFS-associated oncogenesis in nude mouse model. UVRAGFS-NIH3T3 cells from (a) were subcutaneously injected into flanks of nude mice, and tumour growth was measured over time. Circles indicate xenograft tumours at day 38 after inoculation. Data shown are representative of three separate experiments. (e) Immunohistochemistry staining of 3T3-tumours with the indicated antibodies and their quantification in bar graphs (bottom). Arrows denote the mitotic and Ki67+ proliferating cells in the tumour. **P<0.01. Scale bar, 50 μm.

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