Rabbit anti human cd14

The experimental method has been described previously (Feng et al., 2019b (link)). Briefly, an equal amount of total protein was separated by SDS-PAGE gel and then subjected to other immunoblot analysis by antibody mouse anti-human β-actin (Cat No. bsm-33036M, Bioss, Beijing, China), rabbit anti-human E2F1 (Cat No. ab218527, Abcam, Danvers, MA, United States), rabbit anti-human CDK4 (Cat No. ab108357, Abcam, Danvers, MA, United States), rabbit anti-human Cyclin A2 (Cat No. ab181591, Abcam, Danvers, MA, United States), rabbit anti-human Cyclin D3 (Cat No. ab183338, Abcam, Danvers, MA, United States), rabbit anti-human P-Rb (Cat No. ab184796, Abcam, Danvers, MA, United States), rabbit anti-human CD11b (Cat No. ab133357, Abcam, Danvers, MA, United States), rabbit anti-human CD14 (Cat No. ab133335, Abcam, Danvers, MA, United States), and rabbit anti-human EZH2 (Cat No. ab186006, Abcam, Danvers, MA, United States). Immunoreactive bands were then visualized, and the optical densities were measured.

We first used 8% SDS-PAGE to isolate the protein lysate, and then transferred the protein to 0.22-µm PDVF membrane. PVDF membrane was incubated with the corresponding primary antibody at 4 °C overnight and then incubated with horseradish peroxidase-labeled secondary antibody at room temperature. All the experiments were repeated three times. β-actin (Cat No. bsm-33036M, Bioss, Beijing, China), rabbit anti-human LATS2 (Cat No. 5888, CST, Boston, USA), rabbit anti-human CDK4 (Cat No. ab108357, Abcam, Danvers, MA, USA), rabbit anti-human Cyclin A2 (Cat No. ab181591, Abcam, Danvers, MA, USA), rabbit anti-human Cyclin D3 (Cat No. ab183338, Abcam, Danvers, MA, USA), rabbit anti-human P-Rb (Cat No. ab184796, Abcam, Danvers, MA, USA), rabbit anti-human CD11b (Cat No. ab133357, Abcam, Danvers, MA, USA), rabbit anti-human CD14 (Cat No. ab133335, Abcam, Danvers, MA, USA), and rabbit anti-human EZH2 (Cat No. ab186006, Abcam, Danvers, MA, USA).

sGMSCs were respectively labeled with rabbit anti-human CD14, CD31, CD34, CD45, CD73, CD90, and CD105 (Abcam) according to the manufacturer’s instructions. A rabbit anti-human IgG antibody (Abcam) served as a negative control. The tube was incubated in the dark at room temperature (RT) for 2 h. After incubation, cells were washed three times with PBS to remove unbound antibodies. Subsequently, the cells were resuspended and analyzed using a Beckman CytoFLEX flow cytometer (Beckman Coulter, IN, USA) with appropriate settings and compensation adjustments. The negative control was utilized to establish the negative boundary, while the fluorescence intensity and number of positive cells were measured.