Journal: DNA and Cell Biology

Article Title: microRNA133a Targets Foxl2 and Promotes Differentiation of C2C12 into Myogenic Progenitor Cells

doi: 10.1089/dna.2014.2522

Figure Lengend Snippet: Primers Used for Real-Time Polymerase Chain Reaction

Article Snippet: FOXL2 antibody was from Boster Biotechnology.

Techniques: Sequencing

Journal: DNA and Cell Biology

Article Title: microRNA133a Targets Foxl2 and Promotes Differentiation of C2C12 into Myogenic Progenitor Cells

doi: 10.1089/dna.2014.2522

Figure Lengend Snippet: Foxl2 is a direct target of miR-133a. (A) Predicted miR-133a binding sites in Foxl2 3′UTR. Specific locations of the binding sites were marked with red color, and Foxl2 3′UTR was marked with blue color. (B) Alignment between the predicted miR-133a target sites and miR-133a is shown. The conserved, 7-bp seed sequence for miR-133a:mRNA pairing is also indicated. (C) Diagram depicting the pMIR luciferase reporter constructs, containing a cytomegalovirus (CMV) promoter, which was utilized to verify the putative miR-133a binding sites. Luc, luciferase; poly A, poly(A) tail. (D) HEK293 cells were cotransfected with miR-133a, pLuc-FOXL2 3′UTR, along with a pRL-TK reporter plasmid. After 24 h, the luciferase activity was measured. Values are presented as relative luciferase activity after normalization to Renilla luciferase activity. The data are expressed as the mean value±S.E. (error bars) of the results obtained from three independent experiments. *Differences in luciferase activity between miR-133a and negative control transfected cells were significant, p<0.05. (E) Foxl2 expression levels in C2C12 cells after transfection with miR-133a were determined by western blot analysis. As compared with the NC miRNA, miR-133a expression reduced the levels of Foxl2 in C2C12 cells. GAPDH was used as an internal control. Foxl2, forkhead transcriptional factor 2; UTR, untranslated region.

Article Snippet: FOXL2 antibody was from Boster Biotechnology.

Techniques: Binding Assay, Sequencing, Luciferase, Construct, Plasmid Preparation, Activity Assay, Negative Control, Transfection, Expressing, Western Blot, Control

Journal: DNA and Cell Biology

Article Title: microRNA133a Targets Foxl2 and Promotes Differentiation of C2C12 into Myogenic Progenitor Cells

doi: 10.1089/dna.2014.2522

Figure Lengend Snippet: Expression of miRNAs and Foxl2 after using 2% HS differentiation of C2C12 cells for 5 days and Foxl2 knockdown by siRNA. (A) Quantitative analysis of miR-1, miR-133a, and miR-206 by real-time PCR. (B) Quantitative analysis of Foxl2 by real-time PCR. (C) Foxl2 protein expression. (D) Quantitative analysis of Foxl2 by real-time PCR. (E) Quantitative analysis of MyoD, MyHC, MyoG, Caldesmon, and MAML1 by real-time PCR. **p<0.01, *p<0.05.

Article Snippet: FOXL2 antibody was from Boster Biotechnology.

Techniques: Expressing, Knockdown, Real-time Polymerase Chain Reaction