Fig. 7
uc.339 increases the expression of CCNE2 by sequestering its targeting miRNAs. a Immunoblotting for CCNE2 and Vinculin in A549, H460, H1299, and LoVo cells infected with LV-uc.339 or LV-E. b Immunoblotting for CCNE2 and Vinculin in A549, H460, H1299, and LoVo cells transfected with si-uc.339(1) or si-SCR for 72 h. c Immunoblotting for CCNE2 and Vinculin in A549, H460, H1299, and LoVo cells infected with LV-uc.339 and transfected with miR-339, -663b, and -95 or a scrambled miRNA (SCR) for 48 h from transfection. d Representative image of RNA fluorescent in situ hybridization for uc.339, miR-339, miR-663b, and CCNE2 in two nude mice injected subcutaneously with A549 LV-E or A549 LV-uc.339. Scale bar = 200 μm. e Cell viability assay in A549 cells transfected for 72 h with a plasmid-expressing wild-type uc.339 (uc.339 wt) or expressing uc.339 in which the miRNA-binding elements of miR-339, -663b, and -95 have been deleted (uc.339Δ339, uc.339Δ663b, and uc.339Δ95) or with its empty plasmid counterpart. Data presented as mean ± s.d. of experimental triplicates normalized to empty. *P<0.05. Immunoblotting for CCNE2 and Vinculin of the same experiment are shown in the lower panel. f Cell viability assay in A549 LV-uc.339 transfected with si-miR-339, -663b, and -95 or si-SCR for 48 h. Data presented as mean ± s.d. of experimental triplicates normalized to si-SCR. *P<0.05. Immunoblotting for CCNE2 and Vinculin of the same experiment are shown in the lower panel. g Cell viability assay in A549 LV-uc.339 transfected with miR-339, -663b, and -95 or SCR for 48 h. Data presented as mean ± s.d. of experimental triplicates normalized to SCR. *P<0.05. Immunoblotting for CCNE2 and Vinculin of the same experiment are shown in the lower panel. h Cell viability assay in A549 LV-E and A549 LV-uc.339 transfected with two different siRNAs against CCNE2 (si-CCNE2 (1), si-CCNE2 (2)) or si-SCR for 72 h. Data presented as mean ± s.d. of experimental triplicates normalized to LV-E. *P<0.05. Immunoblotting for CCNE2 and Vinculin of the same experiment are shown in the lower panel. The numbers above the bands represent the quantification of the band intensity normalized to Vinculin and to controls
