Cancer cachexia is regulated by selective targeting of skeletal muscle gene products
J. Clin. Invest. Swarnali Acharyya, et al. 114:370 doi:10.1172/JCI20174 [Go to this article.]

Figure 2
Chronic TNF/IFN signaling is required to induce the temporal downregulation of MyHC mRNA. (A) Myotubes were treated with DM containing increasing doses of TNF (5 ng/ml, lane 1; 10 ng/ml, lane 2), IFN (50 U/ml, lane 3; 100 U/ml, lane 4), or TNF plus IFN (5 ng/ml + 50 U/ml, lane 5; 10 ng/ml + 100 U/ml, lane 6). After 48 hours, RNA was prepared and RT-PCR was performed (MyHC IIb; Tn3, troponin T3; α-TM, α-tropomyosin; α-actin). (B) Myotubes were either untreated or treated with TNF (5 ng/ml), IFN (50 U/ml), or TNF plus IFN (5 ng/ml + 50 U/ml) for 48 hours. RNA was prepared, and Northern blots probing for MyHC IIa and IIb were performed. GAPDH was used as a loading control. Numbers are densitometric values normalized to GAPDH. (C) Myotubes were treated with TNF/IFN, and at the indicated times RNA was prepared and Northern blots probing for MyHC type IIa and IIb were performed. (D) Myotubes were treated with TNF/IFN for 24 hours, at which time cells continued to be treated with cytokines or were washed with PBS and switched to DM (Wash). RNA was prepared at the indicated time points, and Northern blots probing for MyHC IIb were performed. Similar results were obtained from two additional independent experiments.