Figure 5. MyBP-C and MyLCs are lost... | Rockefeller University Press

Figure 5.

$Figure 5. MyBP-C and MyLCs are lost selectively during atrophy and are efficiently ubiquitylated by MuRF1 in isolated myofibrils. (A) Equal amounts of the myofibrillar fraction from innervated gastrocnemius muscles and ones denervated for 3 or 10 d from WT mice were analyzed by SDS-PAGE and Coomassie blue staining (left). The intensity of specific Coomassie blue–stained bands was measured, and the ratio to actin was plotted. Error bars represent SEM, n = 5. *, P < 0.05. Shown on the right is Western blot analysis of myofibrillar fraction (0.5 µg, to detect MyLC1) and cytosolic fractions (20 µg, to detect MuRF1) from innervated and denervated muscles from WT mice. (B) MyBP-C and MyLCs are ubiquitylated by purified MuRF1 in isolated myofibrils from control muscles and ones denervated for 3 or 10 d using UbcH1 and His-tagged ubiquitin. His-Ubiquitylated proteins were purified with a nickel column and detected by immunoblotting with specific antibodies.$

MyBP-C and MyLCs are lost selectively during atrophy and are efficiently ubiquitylated by MuRF1 in isolated myofibrils. (A) Equal amounts of the myofibrillar fraction from innervated gastrocnemius muscles and ones denervated for 3 or 10 d from WT mice were analyzed by SDS-PAGE and Coomassie blue staining (left). The intensity of specific Coomassie blue–stained bands was measured, and the ratio to actin was plotted. Error bars represent SEM, n = 5. *, P < 0.05. Shown on the right is Western blot analysis of myofibrillar fraction (0.5 µg, to detect MyLC1) and cytosolic fractions (20 µg, to detect MuRF1) from innervated and denervated muscles from WT mice. (B) MyBP-C and MyLCs are ubiquitylated by purified MuRF1 in isolated myofibrils from control muscles and ones denervated for 3 or 10 d using UbcH1 and His-tagged ubiquitin. His-Ubiquitylated proteins were purified with a nickel column and detected by immunoblotting with specific antibodies.