Journal: Journal of Biological Chemistry

Article Title: Selective Roles for Tumor Necrosis Factor α-converting Enzyme/ADAM17 in the Shedding of the Epidermal Growth Factor Receptor Ligand Family

doi: 10.1074/jbc.m312141200

Figure Lengend Snippet: FIG. 3. Cleavage of EGF ligand ectodomains. Ectodomains were prepared as described under “Experimental Procedures” and incubated in the presence and absence of recombinant human TACE (150 g/ml: AR, HB-EGF; 300 g/ml: BTC, EPR) for 0–24 h. pro-HB-EGFecto was also incubated with recombinant human ADAM10 (150 g/ml) for 0–24 h. Reactions were stopped by the addition of SDS-PAGE sample buffer and analyzed by Western blot analysis using the HA and FLAG antibodies.

Article Snippet: In Vitro Digests—Aliquots (1–10 g) of ectodomain proteins were incubated with 150–300 g/ml recombinant human TACE or ADAM9 or ADAM10 (R & D Systems) or 10 mM Tris, pH 8.0 (as indicated) (31), for 1 or 24 h. Reactions were stopped by adding SDS-PAGE sample buffer, and products were separated by SDS-PAGE, transferred to Immobilon polyvinylidene difluoride, and probed with the indicated antibodies.

Techniques: Incubation, Recombinant, SDS Page, Western Blot

Journal: Cells

Article Title: Ancient Herbal Formula Mahuang Lianqiao Chixiaodou Decoction Protects Acute and Acute-on-Chronic Liver Failure via Inhibiting von Willebrand Factor Signaling.

doi: 10.3390/cells11213368

Figure Lengend Snippet: Figure 6. The therapeutic effect of MHLQD could not be replicated in VWF inhibitor-treated mice. The mice were intraperitoneally injected with D−GalN and LPS to establish ACLF and ALF models. The rADAMTS13 (200 ug/kg), a VWF clearer, was injected into mice 0.5 h after D−GalN/LPS treatment. MHLQD or PBS was orally administrated to mice 1.5 h before and 1.5 h after LPS/D−GalN injection. (A): The experimental design of ACLF mice. (B): The H&E staining of the liver and the liver function levels (ALT and AST) in ACLF mice (n = 5). Scale bar: 0.25 mm. (C): The TUNEL staining and p-MLKL staining in liver tissues of ACLF mice. Scale bar: 0.25 mm (D): The experimental design of ALF mice. (E): The H&E staining of the liver and the liver function levels (ALT and AST) in ALF mice (n = 5). Scale bar: 0.25 mm. (F): The TUNEL staining and p-MLKL staining in liver tissues of ALF mice. Scale bar: 0.25 mm. (G): The protein expression of Caspase 3 and P-MLKL/MLKL in liver tissues of ACLF and ALF mice. ns: no significant. Abbreviation: MHLQD: Ma-Huang-LianQiao- ChiXiaoDou decoction, VWF: von Willebrand factor, ACLF: acute-on-chronic liver failure, ALF: acute hepatic failure.

Article Snippet: The following chemicals were used: LPS (L2630, Sigma-Aldrich, St. Louis, MO, USA); D−GalN (MB1931, Meilunbio, Dalian, China); CCl4 (C805329, Macklin, Shanghai, China); Recombinant ADAMTS13 (4245-AD, R&D systems, Minneapolis, MN, USA).

Techniques: Injection, Staining, TUNEL Assay, Expressing