Fig. 5: Activation of Casp-3 was required for TNF-α-induced pyroptosis in C2C12 myotubes.
From: TNF-α contributes to sarcopenia through caspase-8/caspase-3/GSDME-mediated pyroptosis
A immunoblots of Casp-3 and Cl-Casp-3 in myotubes treated with different concentrations (0, 1, 10, or 100 ng/ml) of TNF-α for 72 h. Relative expression levels of Cl-Casp-3 normalized to β-Tubulin based on densitometric analysis of immunoblots. B–E myotubes were pretreated with Z-DEVD-FMK (50 μM) for 1 h followed by treatment with TNF-α (100 ng/ml) for 72 h. The expressions of GSDME and GSDME-N in myotubes were immunoblotted and analyzed based on densitometric analysis of immunoblots (B). Cell death was determined by measuring LDH release into the cell culture supernatant (C) and staining with Hoechst 33342/PI (D) after myotubes treatments. Scale bar: 50 μm. E Quantitation of positive ratio of PI. Data are expressed as mean ± SD. **P < 0.01, ***P < 0.001, ns: no significance. n = 5 independent experiments.
