Fig. 3: BARD1 is a direct target of miR-210-3p and downregulated in ectopic lesions. | Cell Death & Disease

Fig. 3: BARD1 is a direct target of miR-210-3p and downregulated in ectopic lesions.

From: MiR-210-3p protects endometriotic cells from oxidative stress-induced cell cycle arrest by targeting BARD1

Fig. 3

a qRT-PCR assay of BARD1 mRNA expression in endometrial stromal cells (ESCs) (left panel, n = 5) and Ishikawa cells (right panel) infected with LV-CN, LV-210, LV-In-CN or LV-In-210 under normoxia. The bars show the mean value of three independent experiments ± standard deviation. *P < 0.05, LV-210 versus LV-CN; #P < 0.05, LV-In-210 versus LV-In-CN; paired Studentʼs t-test. b Western blot of BARD1 and BRCA1 protein levels in ESCs (n = 5) and Ishikawa cells infected with LV-CN, LV-210, LV-In-CN or LV-In-210 under normoxia. β-Actin served as the internal control. c Diagram depicting the putative miR-210-3p-binding site within the 3′-UTR of BARD1 and the mutant 3′-UTR of BARD1. d 293 T cells were transiently transfected with a luciferase reporter containing wild type (WT) or mutant (MUT) BARD1 3′-UTR, along with miR-210-3p mimic (miR-210 Mimic) or a negative control (NC) (n = 3). *P < 0.05 versus treatment of NC with WT 3′-UTR; #P < 0.05 versus treatment of miR-210 mimic with MUT 3′-UTR; one-way ANOVA with LSD for multiple comparisons. e qRT-PCR assay of BARD1 mRNA expression in normal endometrium and paired eutopic and ectopic tissues (n = 15). *P < 0.05 versus normal endometria; #P < 0.05 versus eutopic endometria; Mann–Whitney U test. f Scatter diagram showing linear regression and significant Pearson correlation of BARD1 with miR-210-3p in ectopic lesions based on qRT-PCR results (n = 15). P < 0.05. g Immunohistochemistry photomicrographs of BARD1 and BRCA1 in normal endometrium and paired eutopic and ectopic tissues. Scale bars = 50 µm. Original magnification: ×400. h Expressions of BARD1 and BRCA1 in ESCs and endometrial glandular cells by H-score (y-axis) were significantly higher in normal endometria than in eutopic endometria and ectopic lesions, with ectopic lesions showing the lowest staining (n = 27 for normal endometria, n = 57 for eutopic endometria, n = 57 for ectopic lesions). *P < 0.05 versus normal endometria, #P < 0.05 versus eutopic endometria, one-way ANOVA with LSD for multiple comparisons

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