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Fig. 2. MiR-103-3p suppresses TWF1 expression by targeting TWF1 3’UTR. (A) The tentative miR-103-3p (miR-103) target site of the TWF1 3’UTR. (B) Schematic diagram of wild-type TWF1 3’UTR segment (TWF1wt) and mutant TWF1 3’UTR segment (TWF1mut) containing miR-103 target sites cloned in pmirGLO dual-luciferase reporter plasmids and the seed sequence of miR-103 target sites on the TWF1wt. (C) C2C12 cells were co-transfected with scRNA control or miR-103-3p mimic (miR-103) and either TWF1wt or TWF1mut. Luciferase activities were determined 24 h after transfection. (D) The protein levels of TWF1 in C2C12 cells were detected by immunoblotting after 24 h of transfection with scRNA control, miR-103-3p mimic (miR-103), or antimiR-103. The immunoblot intensities were normalized to the amount of β-Actin. The values are shown as relative ratios, with the intensity of the normalized scRNA control set to one. All data are shown as the mean ± SEM (n > 3), where the level of significance is represented as **p < 0.01; ***p < 0.001 vs. scRNA. TWF1, Twinfilin-1; CDS, coding sequence; ns, no significance.
Korean J Physiol Pharmacol 2023;27:277-287
© Korean J Physiol Pharmacol