mf20 rrid ab 2147781  (Developmental Studies Hybridoma Bank)

Journal: bioRxiv

Article Title: NRMLncR, a myocyte-enriched long non-coding RNA, enhances myogenesis in mouse

doi: 10.64898/2026.02.11.704964

Figure Lengend Snippet: (A) Schematic illustration of two shRNA designs targeting NRMLncR-v1 and total NRMLncR-v1/2 for generating stable NRMLncR knockdown C2C12 cell lines. (B) Knockdown efficiency of NRMLncR-v1 and NRMLncR-v1/2 in stable NRMLncR knockdown C2C12 cell lines. n=3. (C) Relative mRNA expression levels in stable NRMLncR knockdown C2C12 cell lines. n=3. (D) Cell proliferation assay in control and NRMLncR knockdown C2C12 cell lines. n=4. (E) Immunofluorescence staining of MyoG and MF20 in control and NRMLncR knockdown C2C12 cultures after 4 days of differentiation. Scale bar: 50 μm. (F and G) Quantification of the percentage of MyoG + nuclei (F) and myonuclei number in MF20 + myotubes (G) in control and NRMLncR knockdown C2C12 cultures after 4 days of differentiation. n=3 for F, n=5 for G. (H) Western blot analysis of myogenic proteins in control and NRMLncR knockdown C2C12 cultures after 4 days of differentiation. (I) Relative mRNA expression of myogenic genes in C2C12 cultures after 4 days of differentiation. n=4. (J) Immunofluorescence staining of MF20 in lentiviral-infected primary myoblasts after 1 and 4 days of differentiation. Scale bar: 50 μm. (K) Relative mRNA expression of myogenic genes in lentiviral-infected primary myoblasts after 1 and 4 days of differentiation. n=3. Data represent mean ± SEM. One- or Two-way ANOVA multiple comparisons test: * p < .05; ** p < .01; and *** p < .001.

Article Snippet: Samples were incubated with primary antibodies at 4 °C overnight, followed by incubation with appropriate fluorescent secondary antibodies and DAPI at room temperature for 1 h. The following primary and secondary antibodies were used: anti-MyoG (DSHB, cat#F5D), MF20 (DSHB, cat#MF-20), FLAG (Sigma-Aldrich, cat#F1804), Alexa 568 goat anti-mouse IgG1 (Invitrogen, cat#A-21124), and Alexa 647 goat anti-mouse IgG2b (Invitrogen, cat#A-21242).

Techniques: shRNA, Knockdown, Expressing, Proliferation Assay, Control, Immunofluorescence, Staining, Western Blot, Infection