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. Author manuscript; available in PMC: 2021 Mar 23.
Published in final edited form as: Dev Cell. 2020 Feb 27;52(6):748–763.e6. doi: 10.1016/j.devcel.2020.01.037

Figure 2. Reduced expression of miRNAs de-represses RBFOX2 in DM1 cardiac cultures.

Figure 2.

(A) Schematic of putative miRNAs targeting Rbfox2 3’-UTR that are downregulated in the hearts of DM1 patients (Kalsotra et al., 2014). (B) Relative expression of indicated miRNAs in HL-1 cells following transfection with DT0 or DT960 plasmids for 48h. n=6 independent transfections. (C) Immunoblot analysis of RBFOX2 protein in HL-1 cells following treatment with scrambled control or indicated miRNA mimics. n=4 independent transfections. (D) Relative luciferase activity derived from Rbfox2 3’-UTR reporter transfected in HEK 293T cells following treatment with scrambled control or indicated miRNA mimics. Direct binding of miRNAs to the seed sequences in Rbfox2 3’-UTR was examined by co-transfecting indicated miRNA mimics and reporters with mutations that would disrupt the predicted miRNA interactions. n=4 independent transfections. (E) Relative luciferase activity derived from Rbfox2 3’-UTR reporter in HEK 293T cells, and (F) Immunoblot analysis of RBFOX2 protein in HL-1 cells after co-transfection with DT0 or DT960 plasmids, and scrambled control or a cocktail of indicated miRNA mimics. n=4 independent transfections. Data are mean ± s.d., and p-values were derived from a parametric t-test (two-sided, unpaired), with Welch’s correction.