Journal: Cancer Management and Research
Article Title: Long Non-Coding RNA XIST Promotes Wilms Tumor Progression Through the miR-194-5p/YAP Axis
doi: 10.2147/CMAR.S297842
Figure Lengend Snippet: miR-194-5p can bind to both XIST and YAP in WT tissues. XIST lncRNA regulates WT progression through the miR-194-5p/YAP axis. ( A ) XIST 3ʹ-UTR wild-type (XIST-wt) sequence containing the miR-194-5p binding site and sequence of the mutant (XIST-mut) miR-194-5p binding site. ( B and C ) Luciferase reporter gene assay (images and histograms) showed lower luciferase activity for miR-194-5p and XIST-wt than XIST-mut ( P < 0.05). TRAF6 was used as an internal control to verify the integrity of the luciferase gene reporter assay. ( D ) YAP 3ʹ-UTR wild-type (YAP-wt) sequence containing the miR-194-5p binding site and sequence of the mutant (YAP-mut) miR-194-5p binding site. ( E and F ) Luciferase reporter gene assay (images and histograms) showed lower luciferase activity for miR-194-5p and YAP-wt than YAP-mut ( P < 0.05). TRAF6 was used as an internal control to verify the integrity of the luciferase gene reporter assay. ( G ) XIST lncRNA expression and ( H ) miR-194-5p in WT G401 cells and normal renal epithelial HK2 cells. ( I ) RT-qPCR analysis of miR-194-5p after transfection of lentiviral XIST, NC, and sh-XIST in WT G401 cells. ( J ) Western blot analysis showed that YAP protein expression can be regulated by miR-194-5p and XIST. One-way ANOVA or two-tailed t -test was performed for comparisons between the two groups. * P < 0.05, *** P < 0.001.
Article Snippet: ChIP was next performed using the human lncRNA array v4 lab-on-a-chip kit (CapitalBio Technology, Beijing, China) according to the manufacturer’s instructions, and the results were analyzed using the GeneSpring software V13.0 (Agilent).
Techniques: Sequencing, Binding Assay, Mutagenesis, Luciferase, Reporter Gene Assay, Activity Assay, Control, Reporter Assay, Expressing, Quantitative RT-PCR, Transfection, Western Blot, Two Tailed Test