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. 2020 Jul 13;11(7):531. doi: 10.1038/s41419-020-2735-2

Fig. 3. GAS5 physically associates with miR-21.

Fig. 3

a Fractionation of HeLa and SiHa cells followed by qRT-PCR. β-Actin served as the cytoplasmic mRNA control. MALAT1 served as the nuclear RNA control. The results showed that GAS5 was mainly located in the cytoplasm. b Bioinformatics manners predicted the potential binding sites within GAS5 and miR-21. c RNA secondary structure of GAS5 predicted by the Vienna RNAfold server. d Compared to co-transfection of the GAS5 plasmid and miR-21 MUT plasmid, co-transfection of the GAS5 plasmid and miR-21 WT plasmid strongly decreased luciferase activity. e RIP assay showed that miR-21 and GAS5 or PDCD4 expression were significantly enriched in the Ago2 pellet compared to the IgG control. f An RNA pull-down assay followed by qRT-PCR was performed to assess the interaction between GAS5 and miR-21 using biotin-labelled GAS5. The miR-21 level in the biotin-GAS5 group was compared to that in the biotin-NC group. U6 was used as the non-specific control. g Rescue experiments showed that the miR-21 mimic could rescue GAS5 overexpression, and that GAS5 also downregulated the expression of miR-21. The data are shown as the means ± SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001.