Journal: Oxidative Medicine and Cellular Longevity

Article Title: Myostatin Activates the Ubiquitin-Proteasome and Autophagy-Lysosome Systems Contributing to Muscle Wasting in Chronic Kidney Disease

doi: 10.1155/2015/684965

Figure Lengend Snippet: TNF- α regulates myostatin expression via nuclear factor (NF)- κ B. (a) The expression of TNF- α mRNA was measured by RT-PCR in rat muscle and is presented as corrected for GAPDH and normalized to Sham group. (b) The expression of TNF-R1 protein was measured by immunoblotting in rat muscle and is presented as corrected for GAPDH and normalized to Sham group. (c) The expression of p-IkBa protein was measured by immunoblotting in rat muscle and is presented as corrected for GAPDH and normalized to sham group. (d) C2C12 cells were infected with a NF- κ B-promoter luciferase construct. Following their differentiation, the myotubes were treated with 100 ng/mL TNF- α . Activation of the NF- κ B promoter at times listed was measured and the fold change over 0 h was quantified. (e) Myotubes were treated with 100 ng/mL TNF- α with or without the NF- κ B inhibitor (QNZ). Representative immunoblots of myostatin and GAPDH and the ratio of myostatin and GAPDH normalized to control. (f) Upper: representative fluorescent images of MHC. Lower: measurements of average myotube diameter after 24 hr of 100 ng/mL TNF- α treatment with or without the QNZ. Scale bar = 50 μ m. Values were described means, with SD represented by vertical bars. Significantly different ( ∗ P < 0.05, ∗∗ P < 0.01) from control group, ( † P < 0.05, n = 3 independent experiments) from TNF- α group.

Article Snippet: Vectastain Elite ABC Peroxidase kit and Imm PACT DAB peroxidase substrate were from (Vector Laboratories, Burlingame, CA, USA), TNF- α was from R&D Systems (Minneapolis, MN), QNZ, the NF- κ B inhibitor, was from Enzo Life Sciences (Ann Arbor, MI), and the myostatin siRNA and control siRNA were from Santa Cruz Biotechnology.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Infection, Luciferase, Construct, Activation Assay