Fig. 8

Integrity of the circNTNG1/miR-19b-3p/HOXA5 regulatory axis. a. Immunoblotting of HOXA5 and Slug in 769P cells circNTNG1 overexpression salvage experiment. Cells were treated with circNTNG1 overexpression with/without miR-19b-3p mimics, and further salvaged with HOXA5 overexpression. GAPDH was used as positive control. b. Representative images (left) and quantification (right) data of Transwell migration/invasion assay of 769P cells circNTNG1 overexpression salvage experiment. c. Immunoblotting of HOXA5 and Slug in Caki-1 cells circNTNG1 overexpression salvage experiment. Cells were treated with circNTNG1 overexpression with/without miR-19b-3p mimics, and further salvaged with HOXA5 overexpression. GAPDH was used as positive control. d. Representative images (left) and quantification (right) data of Transwell migration/invasion assay of Caki-1 cells circNTNG1 overexpression salvage experiment. e. Representative images (left) and in vivo luciferase activity quantification (right) data of mouse tail-vein lung metastasis model. 769P cells with vector, circNTNG1-overexpression, circNTNG1-overexpression+miR-19b-3p overexpression or circNTNG1-overexpression+miR-19b-3p overexpression+HOXA5 overexpression were used in the injection. Images were taken 8 weeks after the injection. f. Representative HE-stain images (left) and lung-metastasis foci quantification (right) data of 769P mouse tail-vein lung metastasis model. g. Representative IHC images of HOXA5 in lung-metastasis foci from 769P mouse tail-vein lung metastasis model. h. Schematic diagram of the intact circNTNG1/miR-19b-3p/HOXA5 pathway in controlling the Slug expression and EMT activity via methylation change. Data are mean ± SD, n = 3