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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: LINC00673 is activated by YY1 and promotes the proliferation of breast cancer cells via the miR-515-5p/MARK4/Hippo signaling pathway

Fig. 5

LINC00673 regulates MARK4 expression by competing for miR-515-5p. a The subcellular distribution of LINC00673 in MDA-MB-231 and MDA-MB-453 cells. GAPDH was used as the cytoplasmic control, and U1 served as the nuclear control. b-c The relative expression of miRNAs was determined by qRT-PCR after the knockdown of LINC00673. d The expression of miR-515-5p was measured after the knockdown of LINC00673 by using qRT-PCR in MDA-MB-231 and MDA-MB-453 cells. e Expression of LINC00673 in miR-515-5p mimics or inhibitor transfected MDA-MB-231 and MDA-MB-453 cells, as determined by qRT-PCR. f-g Elevation and depression of miR-515-5p was inversely related to MARK4 expression, as determined by qRT-PCR and western blotting. h Luciferase reporter assays were used to verify the targeted binding between LINC00673 or MARK4 3’untranslated region (UTR) and miR-515-5p. i The expression of MARK4, YAP and TAZ in MDA-MB-231 and MDA-MB-453 cells transfected with LINC00673 siRNA or cotransfected with a LINC00673 siRNA and an miR-515-5p inhibitor, as determined by qRT-PCR. j Western blot analysis of MARK4, p-YAP, YAP and TAZ expression levels in MDA-MB-231 and MDA-MB-453 cells transfected with LINC00673 siRNA or cotransfected with LINC00673 siRNA and miR-515-5p inhibitor. k-l CCK-8 assays were used to examine the cell proliferation ability after LINC00673 knockdown in MDA-MB-231 and MDA-MB-453 cells transfected with miR-515-5p inhibitors and LINC00673-overexpressing cells transfected with miR-515-5p mimics. The data are presented as the mean ± the SD of three independent experiments. *P < 0.05, ** P < 0.01, and *** P < 0.001

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