Clone 7h8.2c12

Anti-PARP, anti-β-actin, anti-caspase-3, anti-caspase-9, VDAC, anti-phosphorylated ERK1/2, anti-phosphorylated p38, anti-phosphorylated JNK, anti-phosphorylated IκBα, anti-ERK1/2, anti-p38, anti-JNK, anti-Bcl-2, and anti-IκBα were purchased from Cell Signaling Technology, Inc. (Beverly, MA, USA). α-Tubulin, anti-Bax (6A7) and His-probe were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against cytochrome c (for immunofluorescence, clone 6H2.B4; for western blot analysis, clone 7H8.2C12) were acquired from BD Pharmingen (San Diego, CA, USA), and the anti-cytochrome oxidase subunit IV (COX IV) antibody was purchased from Abcam (Cambridge, UK). Dichlorodihydrofluorescein diacetate (H₂DCFDA), MitoSOX, DAPI, and DiOC₆ were obtained from Molecular Probes (Eugene, OR, USA) and z-VAD-fmk, N-acetyl-cysteine (NAC), horseradish peroxidase-conjugated anti-mouse IgG and anti-rabbit IgG, and specific inhibitors of ERK (U0126), p38 mitogen-activated protein kinase (MAPK; SB203580), JNK (SP600125), and nuclear factor-κB (NF-κB; BAY 11-7082) were purchased from Calbiochem (San Diego, CA, USA). Anti-F4/80 and anti-CD11b were performed as recommended by the manufacturer (eBioscience, San Diego, CA, USA).

ECs were stained with rat monoclonal anti-mouse CD31 antibody (PEACAM-1; clone MEC 13.3, 1:500; BD Pharmingen, Heidelberg, Germany) and detected using an Alexa Fluor 488-conjugated polyclonal goat anti-rat antibody (1:500; Invitrogen). Proliferating ECs in tumor tissue were stained with a rat anti-mouse CD105 (endoglin) PE-conjugated monoclonal antibody (1:500, clone: MJ7/18; eBioscience, Frankfurt am Main, Germany). Perivascular cells were stained with Cy3-conjugated mouse monoclonal anti-α-smooth muscle actin antibody (1:1,000, α-SMA clone 1A4; Sigma-Aldrich, Munich, Germany). Mitochondrial morphology and membrane potential were visualized by MitoTracker (RedCMXRos) staining (Invitrogen). For cytochrome c staining, a mouse monoclonal anti-cytochrome c antibody was used (1:500; clone 7H8.2C12; BD Biosciences) and detected with a secondary Alexa Fluor 488-conjugated polyclonal goat anti-mouse antibody (1:500; Invitrogen). For VE-cadherin staining, a monoclonal rabbit anti-VE-cadherin antibody (clone D87F2; Cell Signaling) was used and detected with a secondary Alexa Fluor 488-conjugated polyclonal goat anti-rabbit antibody (1:500; Invitrogen).