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. 2018 Jul 5;26(6):837–846. doi: 10.3727/096504017X14920318811721

Figure 4.

Figure 4

Regulation of RAP2B by XIST and miR-320b in human OS cells and RAP2B as a direct target of miR-320b. (A) The putative binding sequences of miR-320b and 3′-UTR of RAP2B. (B) The RAP2B-WT/RAP2B-MUT vectors and miR-NC/miR-320b mimics were cotransfected into HEK 293T cells, respectively. Luciferase activity of the RAP2B-WT vector was reduced in cells cotransfected with miR-320b mimics. The repression of luciferase activity by miR-320b was not shown in cells transfected with RAP2B-MUT. (C) Western blot assay showed that the expression of RAP2B was downregulated by miR-320b overexpression in both Saos-2 and MG63 cell lines. (D) Western blot results showed that knockdown of XIST also downregulated RAP2B in both Saos-2 and MG63 cells. (E) RAP2B knockdown was achieved by si-RAP2B as demonstrated by Western blot assay, which showed a much lower protein expression of RAP2B. (F, G) MTT assay results showed that OS cell growth was attenuated in response to RAP2B inhibition by si-RAP2B. Data are presented as mean ± SD of three independent experiments. *p < 0.05 compared to the NC group.