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. 2016 Mar 15;7(17):23197–23211. doi: 10.18632/oncotarget.8099

Figure 5. 3’UTRs and miRNA dependency of reciprocal interaction of ceRNAs.

Figure 5

a. and b. Western blot analysis indicated that compared to the untreated control group, ectopic expression of STARD13 3’UTR upregulated protein levels of STARD13, CDH5, HOXD1, and HOXD10 in both MDA-MB-231 a. and MCF-7 cells b.. c. and d. qRT-PCR analysis indicated that compared to the untreated control group, ectopic expression of STARD13 3’UTR didn’t affect mRNA levels of CDH5, HOXD1, and HOXD10 in both MDA-MB-231 c. and MCF-7 cells d.. e. and f. Western blot analysis revealed that protein levels of STARD13, CDH5, HOXD1, and HOXD10 decreased as a result of transfection with siSTARD13. g. and h. qRT-PCR analysis indicated that compared to the untreated control group, knockdown of STARD13 3’UTR didn’t affect mRNA levels of CDH5, HOXD1, and HOXD10 in both MDA-MB-231 g. and MCF-7 cells h.. i. and j. Protein level of STARD13 increased as a result of ectopic expression of its ceRNAs 3’UTRs (upper panel), while knockdown of its ceRNAs decreased the expression of STARD13 (lower panel) in both MDA-MB-231 i. and MCF-7 cells j.. k. Luciferase activity in MCF-7 cells co-transfected with STARD13 3’UTR and a Luc-ceRNA-3’UTR reporter construct. Ectopic expression of STARD13 3’UTR in Luc-ceRNA-3’UTRs-expressing cells increased luciferase activity compared to the control vector group. l. Luciferase activity in MCF-7 cells co-transfected with siSTARD13 and a Luc-ceRNA-3’UTR reporter construct. siSTARD13-activated knockdown of STARD13 in Luc-ceRNA-3’UTR-expressing cells decreased luciferase activity compared with siRNA NC. However, in the presence of siDicer, the effects were reversed. Data were presented as mean ± s.d.; *P < 0.05, **P < 0.01 vs. Control. (m and n) MDA-MB-231 m. and MCF-7 cells n. were transfected with siSTARD13 or along with siDicer followed by western blot assays. Although knockdown of STARD13 decreased protein levels of CDH5, HOXD1, and HOXD10, siDicer reversed the effects. However, despite in the presence of siDicer, siSTARD13 still reduced protein level of STARD13.