Skip to main content
Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: HES1-mediated down-regulation of miR-138 sustains NOTCH1 activation and promotes proliferation and invasion in renal cell carcinoma

Fig. 5

miR-138-2p directly regulates MAML1, APH1A and NOTCH1 in ccRCC cells. a and b Regulation of MAML1 and APH1A by miR-138-5p. 786-O and CAKI-1 cells were transfected with miR-138-5p mimics, inhibitor or with their corresponding negative controls. After transfection, total RNA and whole cell lysates were prepared and analyzed by qPCR (n = 3) (a) and immunoblotting (b), respectively. β-tubulin was used as a loading control. c and d Regulation of NOTCH1 by miR-138–2-3p. 786-O and CAKI-1 cells were transfected with miR-138-2p mimics, inhibitor or with their corresponding negative controls. After transfection, total RNA and whole cell lysates were prepared and analyzed by qPCR (n = 3) (c) and immunoblotting (d), respectively. β-tubulin was used as a loading control. e Luciferase reporter assay. 786-O (left) and CAK-1 (right) cells were transfected with the luciferase reporters bearing wild-type or mutated 3’-UTR of MAML1, APH1A or NOTCH1 together with the control expression plasmid or with pre-miR-138–2 expression plasmid. After transfection, cells were lysed and their luciferase activities were measured (n = 3). f and g Mouse Xenograft. Nude mice were injected with the parental 786-O (upper) or with 786-O cells stably overexpressing miR-138–2 (lower). The representative tumors were shown (f). Tumor volume and weight were also calculated (n = 5) (g). h Immunoblotting. Whole cell lysates were prepared from the tumor tissues and analyzed for the indicated proteins. β-tubulin was used as a loading control

Back to article page