Journal: Cell Reports

Article Title: Nfix Regulates Temporal Progression of Muscle Regeneration through Modulation of Myostatin Expression

doi: 10.1016/j.celrep.2016.02.014

Figure Lengend Snippet: Nfix Is Expressed by SCs and Its Absence Leads to a Reduced Myofiber Cross Sectional Area and to an Increased Slow MyHC Expression (A) Immunofluorescence analysis of Nfix expression (green) in Pax7 + SCs (red) on a Tibialis anterior muscle section of a Mlc3f-lacz mouse, where myonuclei are in purple (β-gal). Hoechst was used to stain nuclei (n = 4 mice). The scale bar represents 25 μm. (B) H&E staining on Tibialis anterior muscles from WT and Nfix -null mice (n = 5 mice). The scale bar represents 100 μm. (C) Graphical representation of the myofiber cross sectional area (CSA) in WT and Nfix -null muscles. The plotted values represent the distribution of n = 65 measurements on five random microscope fields ( ∗∗∗ p < 0.001 and two-tailed unpaired t test). The data are presented as mean ± whiskers from min to max. (D) Western blot analysis of slow MyHC expression in WT and Nfix -null Soleus (SOL) and EDL muscles. The β-tubulin was used to normalize. See also Figure S1 .

Article Snippet: The following primary antibodies and dilutions were used: rabbit anti-Nfix (1:5,000, Geneka Biotechnology); mouse anti-β-tubulin (1:5,000, Covance); mouse anti-slow MyHC (Bad5, 1:2, DSHB); mouse anti-total MyHC (MF20, 1:5, DSHB); mouse anti-Myogenin (IF5D, 1:3, DSHB); rabbit anti-Myostatin (1:500, Millipore); mouse anti-GAPDH (1:5,000, Sigma-Aldrich); rabbit anti-Smad3 (1:1,000, Abcam); rabbit anti-pSmad3 (1:1,000, Abcam).

Techniques: Expressing, Immunofluorescence, Staining, Microscopy, Two Tailed Test, Western Blot