Fig. 4

miR-19b-3p regulates the progression of RCC through inhibiting expression of HOXA5 a. The binding site and sequence of miR-19b-3p and HOXA5 3′-UTR region. b. Luciferase reporter assay with vector, HOXA5 wild-type sequence and HOXA5 mutant sequence transfected miR-19b-3p inhibitor or miR-19b-3p mimics. Vector group was used as normalization control. c. qRT-PCR quantification of HOXA5 in 769P and Caki-1 cells transfected with control mimics NC/miR-19b-3p mimics. The mimics NC group was used as normalization. d. qRT-PCR quantification of HOXA5 in 769P and Caki-1 cells transfected with control mimics NC/miR-19b-3p inhibitor. The mimics NC group was used as normalization. e. Representative images (left) and quantification (right) data of Transwell migration/invasion assay of 769P cells with vector/HOXA5-overexpression. Cell number was determined by counting five random fields under microscope. f. Proliferative activity of 769P cells with vector/HOXA5-overexpression measured by CCK8 assay. Levels were normalized to day 0. g. Representative images (left) and quantification (right) data of Transwell migration/invasion assay of Caki-1 cells with vector/HOXA5-overexpression. Cell number was determined by counting five random fields under microscope. h. Proliferative activity of Caki-1 cells with vector/HOXA5-overexpression measured by CCK8 assay. i. Representative images (left) and in vivo luciferase activity quantification (right) data of mouse tail-vein lung metastasis model. 769P cells transfected with vector/ HOXA5-overexpression plasmid were used in the injection. Images were taken 8 weeks after the injection. j. Representative HE-stain images (left) and lung-metastasis foci quantification (right) data of 769P mouse tail-vein lung metastasis model. k. Representative IHC images of HOXA5 in lung-metastasis foci from 769P vector/HOXA5-overexpression groups. Data are mean ± SD, n = 3