Skip to main content
. 2016 Jan 13;7(13):15618–15631. doi: 10.18632/oncotarget.6905

Figure 6. The LILRB3 ligand co-purifies with cytokeratin 8 from the lysates of glandular epithelial cell lines.

Figure 6

A. Coomassie blue stained SDS-PAGE gel of proteins immunoprecipitated by LILRB3*12-Fc from membrane-rich lysates of the epithelial cell lines MCF-7, T47D, and HCT116 compared with the non-epithelial cell lines BJAB and Daudi. The high molecular weight molecules (>64 kDa) in lanes 2-7 are LILR-Fc proteins. The Fc negative control protein in lane 1 has an apparent molecular weight of ∼40kDa. B. A repeat Coomassie stained SDS-PAGE gel of proteins immunopreciptated by LILRB3*12-Fc from Daudi and MCF-7 cells. Bands were excised from the gel and were identified by mass spectrometry as cytokeratins (CK) 8, 18 and 19, β-actin and farnesyl diphosphate synthase (FDPS). C. Isolation of ligand of LILRB3*12 from cell lysates with the anti-human cytokeratin 8 specific mAb 1E8: Ligands captured from MCF-7 lysates (but not from Daudi B cells) on 1E8-coated microsphere beads were cultured with 2B4 reporter cells. Beads coated with isotype control antibody (iso) prior to incubation with lysates were used as a negative control. Beads coated in an anti-LILRB3 antibody were used as a positive control. The results shown are from 4 replicate experiments. Bars indicate mean values. Statistical significance was assessed using a two-tailed Mann Whitney test (**p<0.001).