Journal: American Journal of Cancer Research
Article Title: Transcription factor STAT4 counteracts radiotherapy resistance in breast carcinoma cells by activating the MALAT1/miR-21-5p/THRB regulatory network
doi: 10.62347/VSJU7227
Figure Lengend Snippet: Validation of the targeting correlation between MALAT1 and miR-21-5p in breast carcinoma (BC) cells. A: Venn diagram of StarBase, LncBase predicted miRNAs downstream of MALAT1 and BC-associated miRNAs in GeneCards; B: Diagram of the predicted binding loci of MALAT1 and miR-21-5p by StarBase; C: RT-qPCR detection of miR-21-5p evels in BC tissues and adjacent normal counterparts, n = 66, * indicates comparison with normal counterparts adjacent to cancer, *P < 0.05; D: Correlation analysis of miR-21-5p and MALAT1 in 66 BC tissue samples; E: The targeting correlation between MALAT1 and miR-21-5p by dual luciferase reporter assay, *P < 0.05 versus mimic NC group; F: The binding of biotin-labeled miR-21-5p-WT and mutant sequence miR-21-5p-MUT to MALAT1 by RNA-pull-down assay, *P < 0.05; G: The binding of biotin-labeled MALAT1-WT and mutant sequence MALAT1-MUT to miR-21-5p by RNA-pull-down assay, *P < 0.05; H: RT-qPCR to detect MALAT1 levels in MCF-7R, *P < 0.05 versus oe-NC group; I: RIP assay to detect MALAT1 enrichment within the RISC core element Ago2 in MCF-7R, *P < 0.05; J: RIP assay to detect miR-21-5p enrichment within the RISC core element Ago2 in MCF-7R, *P < 0.05; K: RT-qPCR to quantify miR-21-5p in MCF-7R, *P < 0.05.
Article Snippet: Dual-luciferase reporter (DLR) gene assay Reporter plasmids MALAT1-WT and MALAT1-MUT were obtained by cloning the predicted binding site of MALAT1 with miR-21-5p and a mutated MALAT1 fragment into the Pmir-GLO Dual-Luciferase miRNA Target Expression Vector (Promega, USA).
Techniques: Binding Assay, Quantitative RT-PCR, Comparison, Luciferase, Reporter Assay, Labeling, Mutagenesis, Sequencing, Pull Down Assay
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