Journal: bioRxiv

Article Title: Metavinculin mediates differentiation cues into myogenic gene expression

doi: 10.1101/2024.02.21.578728

Figure Lengend Snippet: a Relative expression of canonical Wnt target genes Myc, Ccnd1, Ascl2, Id2, Tcf4 and Mmp9 in C2C12 cells cultured under various conditions. b Western blot showing canonical Wnt mediator Ctnnb (β-catenin) protein level in C2-Scr and C2-shmVcl2 cells cultured as indicated. c Co-immunoprecipitation analysis showing direct vinculin and Ctnnb interaction in C2-Scr and C2-shmVcl2 cells grown under indicated conditions. d Bar graph showing RT-qPCR validation of Wnt target genes activation by canonical Wnt activator BML-284 in C2C12 cells. e Relative expression of myogenic differentiation associated genes in C2C12 cells treated with BML-284 or vehicle in DM media shown in RT-qPCR. f Myogenic protein expression in BML-284 or vehicle treated C2C12 cells cultured under DM conditions is shown in western blot. g Representative images of Myh stain (green) and DAPI (blue) in C2C12 cells treated with BML-284 or vehicle, Scale bar 50 μM. Fusion index is shown in a bar graph. Actb act as a loading control in western blot. Data are shown as mean value ±SEM, n=3. p value * ≤0.05; **≤0.01; ***≤0.001; ****≤0.0001.

Article Snippet: Wnt activator BML-284 (5 mM) (AdooQ, USA) or inhibitor KY02111 (10 mM) (AdooQ, USA) and vehicle was treated in indicated culturing conditions and refreshed during media change.

Techniques: Expressing, Cell Culture, Western Blot, Immunoprecipitation, Quantitative RT-PCR, Activation Assay, Staining