Skip to main content
. 2020 Nov 5;8:552020. doi: 10.3389/fcell.2020.552020

FIGURE 6.

FIGURE 6

LncRNA GAS5 competitively binds miR-23a to upregulate PTEN expression. The downstream regulatory molecules of GAS5 were analyzed and verified. (A) the biological website confirmed that lncRNA GAS5 is located in the cytoplasm; (B) nuclear/cytosol fractionation experiments confirmed the subcellular localization of lncRNA; (C) FISH assay confirmed that lncRNA GAS5 is localized in the cytoplasm; (D) RT-qPCR detection of the expression of mioR-23a in tissue before and after CGA treatment; (E,F) dual luciferase gene reporter assay accurately verified the existence of binding regions of lncRNAGAS5 and miR-23a, and miR-23a and PTEN genes; (G) RNA pull-down assay found there is a binding region between lncRNA GAS5 and miR-23a; (H) RT-qPCR was used to detect the expression of miR-23a and PTEN; (I) Western blot was used to detect the protein level of PTEN. All the experiments were repeated 3 times. The data in (G,I) were analyzed using one-way ANOVA and Tukey’s multiple comparison test; the data in (B,F,H) were analyzed using two-way ANOVA and Tukey’s multiple comparison test; and the data in (D) were analyzed using independent t-test; compared with sh-NC or kd-NC groups, **p < 0.01.