Journal: Cancer immunology research
Article Title: C3aR Signaling Inhibits NK-cell Infiltration into the Tumor Microenvironment in Mouse Models
doi: 10.1158/2326-6066.CIR-21-0435
Figure Lengend Snippet: a. Immunoprecipitation to assess interaction between C3aR and CD11a (αL subunit that is specific to LFA-1)(n=2). b, c. Representative CD11a staining in activated human NK cells with or without C3a stimulation (n=3). d, e. Representative LFA-1 epitope 24 staining in activated human NK cells with or without C3a stimulation (n=3). f, g. Phospo-CD18 (T758) staining with or without C3a stimulation (n=3). Scale bar: 50 µm. Boxed areas indicate area shown in zoomed images.
Article Snippet: Primary rabbit anti-human CD11a (Cat#ab52895, Abcam, USA), primary rabbit anti-human CD18 (phospo-T758; Cat#ab63388, Abcam, USA), primary mouse anti-human CD11a+CD18 antibody [24] (Cat# ab13219, Abcam, USA), or respective control antibody (Rabbit IgG and Mouse IgG1) was diluted as suggested by the provider, infused into the microfluidic devices, and incubated for overnight at 4 o C. Samples were then washed three times and incubated with diluted secondary antibody (1:200 dilution; anti-mouse goat IgG, Cat# A32723 or anti-rabbit goat IgG, Cat# A32721, Thermofisher Scientific, USA) in the dark at room temperature for one hour.
Techniques: Immunoprecipitation, Staining