Anti 15 lox 1

The 15-LOX-1 inhibitor PD 146176 was purchased from Selleckchem (Houston, TX, USA) and Calbiochem (San Diego, CA, USA). Anti-15-LOX-1 was purchased from Abcam (Cambridge, MA, USA). Anti-p53, anti-macroH2A2, and FITC-conjugated Anti-15-LOX-1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-cleaved PARP, anti-cleaved Caspase3, anti-H2AX, and anti-γH2AX were obtained from Cell Signaling (Beverly, MA, USA). Anti-actin was purchased from Sigma (St. Louis, MO, USA).

The colon mucosa or splenic mononuclear cells, hucMSCs, and RAW 264.7 cells were homogenized and modified in RIPA lysis buffer, added with proteinase inhibitors (Vazyme Biotech, Shanghai, China). Protein samples (two hundred micrograms) were separated on a 12% SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Sources and dilution factors of primary antibodies were the following: anti-caspase-3 (1 : 800; Bioworld), anti-PCNA (1 : 1000; Cell Signaling Technology), anti-15-lox-1 (1 : 1000; Abcam), and anti-β-actin (1 : 800; Bioworld). After incubation with the primary antibodies overnight at 4°C, the blots were incubated with the secondary antibodies for 1 h at room temperature and then were visualized by chemiluminescence (Millipore, USA) and detected by using the imaging software (GE Healthcare, Life Sciences, USA).