Journal: Intractable & Rare Diseases Research
Article Title: The novel role of IFITM1-3 in myogenic differentiation of C2C12 cells
doi: 10.5582/irdr.2023.01050
Figure Lengend Snippet: Knockdown of Ifitm1, 2, and 3 by siRNAs blocks myogenic differentiation in C2C12 cells. (A) Microscopic images of Giemsa staining for C2C12 myoblasts on day 3 of myogenic differentiation after transfection with siRNAs targeting Ifitm1-3. Two different siRNAs were used for each targeting gene. Transfection without siRNAs, but with transfection reagent was set as mock. Magnification: 100×. (B) Percentage fusion on day 3 of myogenic induction and transfection with siRNAs as described above, calculated by dividing the number of nuclei within multinucleated myofibers by the total number of nuclei. NC represents the group without siRNAs and transfection reagents. (C) Downregulated protein expression of myogenin, MyoD, Myf5, and desmin after interference by siRNAs targeting Ifitm1-3. (D) Quantification of band intensity as described above is shown. The level of proteins was normalized to that of GAPDH. Statistical significance: *P< 0.05; **P < 0.01; ***P < 0.001; NS, not significant.
Article Snippet: The membranes were blocked with 5% skim milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) for 2 h at room temperature, and then the membranes were incubated with primary antibodies, namely, rabbit anti-desmin (ab32362, Abcam, USA), rabbit anti-MyoD (ab203383, Abcam, USA), rabbit anti-MYF5 (ab125301, Abcam, USA), rabbit anti-IFITM1 (bs-1031R, Bioss, China), rabbit anti-IFITM2 (bs-15517R, Bioss, China), mouse anti-IFITM3 (bsm-51629M, Bioss, China), and rabbit anti-GAPDH (ab8245, Abcam, USA), at 4°C overnight.
Techniques: Staining, Transfection, Expressing