Additional file 9: of Circular RNA hsa_circ_0008305 (circPTK2) inhibits TGF-β-induced epithelial-mesenchymal transition and metastasis by controlling TIF1γ in non-small cell lung cancer

Figure S7. CircPTK2 overexpression attenuates NSCLC cell metastasis in vivo by the bioluminescent imaging. (A) CircPTK2 expression in A549 cells stably overexpressing circPTK2. pLV-Luci(2A)Puro lentiviral expression vector (upper panel) was used to stably overexpress circPTK2. The empty vector was served as negative control. CircPTK2 expression was determined by qRT-PCR (bottom panel). (B) Schematic flowchart of the in vivo metastasis experiments with A549 cells stably transfected with pLV-circPTK2 or vector (i.v.) and TGF-β1 (i.p.) injected into BALB/c nude mice (n = 10 mice per group in circPTK2 + TGF-β1 and vector + TGF-β1). (C) Representative images of in vivo bioluminescence of mice injected with circPTK2-overexpressed A549 cells or vector-control cells at day 42 post-inoculation. Color bar represents extent of luciferase bioluminescence intensity (blue, green and red indicate low, medium and high intensity, respectively). (D) Quantification of radiance emitted from active luciferase in lung of mice (n = 10 mice for each group) at day 42. (E) Representative images showing metastatic nodules established in lung taken from the mice injected with circPTK2-overexpressed A549 cells or vector control cells at day 49 (upper). Scale bar, 4 mm. H&E staining was performed for histological confirmation of metastasizing tumor cells in lung (bottom). Scale bar, 100 μm. Green arrowhead indicate micrometastasis. (F) Gross view of liver of mice at day 49 post-inoculation (upper) and microscopic images of H&E staining for liver metastases (bottom). Scale bar, 4 mm or 100 μm. Green arrowhead indicate micrometastasis. (G) Dot plots showing the distribution of the number of micrometastases in per section of liver (n = 10 mice for each group). *P < 0.05; **P < 0.01; ***P < 0.001. (TIF 5869 kb