Fig. 4

NORAD increases the expression of RhoA and acts as a ceRNA for miR-125a-3p. a RhoA expression is negatively correlated with miR-125a-3p in GSE32688. The results were expressed as RMA Values; b NORAD and RhoA mRNA levels are positively associated in 33 PDAC tissues; c miR-125a-3p can complement the seed region of RhoA, which is similar to the NORAD MRE. d NORAD increases RhoA protein level. Upper panel: western blot of RhoA was performed. Relative density was shown below each lane. Bottom panel: IHC staining of orthotopic implantation tumor tissues; e RhoA protein level is increased by NORAD-wt but not by NORAD-mut; f The 3′-UTR of RhoA (wt or mut) was fused to the luciferase coding region and transfected into SW1990 with along with a miR-125a-3p mimic to confirm that RhoA is a target of miR-125a-3p. A miR-239b-5p mimic was used as a negative control; g RhoA-3′-UTR-wt-luc was transfected into SW1990-sh-NC or SW1990-sh-NORAD. Inhibitors of miR-125a-3p or miR-239b-5p were added. Rno-miR-239b-5p was used as a negative control. The histogram indicates the luciferase values measured 48 h after transfection. *, statistical significance, P < 0.05, **p < 0.01, ***p < 0.001