Pe conjugated cd11b antibody

For detection of the cell differentiation antigen, CD11 antigen-like family member B was used (CD11b), after 72 h of treatment, the cells were collected (1x10 6 /group) and washed with three times with pre-cooled PBS, then incubated with phycoerythrin (PE) conjugated CD11b antibody (12011342; eBioscience, Inc., San Diego, CA, USA) at 4˚C for 30 min in the dark (32) . The cells were then analyzed using flow cytometry (BD FACSVantage; BD Biosciences, San Jose, CA, USA) and CellQuest Pro software version 5.1 (BD Biosciences).
Indirect immunofluorescence assay. Cells were fixed with 4% paraformaldehyde for 20 min, subsequently, permeabilized with 0.1% Triton X-100 (in PBS) for 15 min, and then blocked in 10% goat serum (in PBS) for 30 min at room temperature. Slides were then incubated overnight with the indicated primary antibodies. Secondary goat antibody against rabbit-IgG-TRITC (1:200; Beijing Zhongshan Golden Bridge Biotechnology) was used to detect rabbit IgG for 1 h at room temperature. The nuclei were stained using DAPI at room temperature. Finally, coverslips were immobilized by 70% glycerol and viewed under a fluorescence microscope (Nikon, Tokyo, Japan).
Statistical analysis. All data were performed using the SPSS 17.0 software (SPSS, Inc., Chicago, IL, USA). Results are represented as the mean ± SD. The Student's t-test was used for statistical analysis.