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. 2016 Aug 3;113(33):E4828–E4836. doi: 10.1073/pnas.1609792113

Fig. 4.

Fig. 4.

The gene encoding the Homeobox protein SIX4 is a direct target of miR-309. (A) Venn plot shows the overlapping up-regulated DEGs after miR-309 knockdown with in silico-predicted gene targets by using five computational programs. (B) The putative miR-309 binding site in 3′-UTR of AAEL010327 is predicted by using software. (C) The expression profile of the SIX4 mRNA during ovarian development in the mosquito. (D) Luciferase reporter assay showing that miR-309 directly degrades the 3′-UTR of AAEL010327 in vitro. (E) Immunoprecipitation (RIP) assay demonstrating that miR-309 directly targets SIX4 in vivo. The SIX4 mRNA was significantly enriched after immunoprecipitation with antibodies against the Anti-Ago. Data represent the fold change to the IgG-precipitated mRNA with three samples, and each sample in triplicate. The error bars shown in C–E indicate the mean ± SEM, **P < 0.01, ***P < 0.001.