Figure 2.

LINC00461 up‐regulates HOXA10 by binding to miR‐195. A, LINC00461 is presumed to bind to the 3'untranslated regions (3'UTR) of miR‐195 using RNA22. B, A combination of miR‐195 mimic and pLINC00461‐Wt significantly reduces the luciferase activity, suggesting that LINC00461 binds to the 3'UTR of miR‐195 (*P < .05, vs the control group). C, miR‐195 is presumed to bind to the 3'untranslated regions (3'UTR) of HOXA10 using RNA22; D, a combination of miR‐195 mimic and pHOXA10‐Wt significantly reduces the luciferase activity, suggesting that LINC00461 binds to the 3'UTR of HOXA10 (*P < .05, vs the control group); E, LINC00461 is predominantly localized in the cytoplasm observed by FISH test. F, Ago2 expression is detected by Western blot analysis (*P < .05, vs the IgG group). G, LINC00461 directly bound to Ago2 by RIP assay. H, LINC00461 binds to miR‐195, determined by RNA pull‐down test (*P < .05, vs the miR‐195‐Wt group). The results of luciferase activity, Western blot analysis and RNA pull‐down test were expressed as mean value ± standard deviation. Comparisons between two groups were conducted by t test. The experiment was run in triplicate independently. The comparisons among three groups were conducted by one‐way ANOVA. miR‐195, microRNA‐195; ANOVA, analysis of variance; Wt, wild‐type; HOXA10, Homeobox A10; and Mut, mutant type