Figure 1. Validation of chemokine-specific pAb use for FC.

HEK 293T cells were transfected (Tx) with individual pIRES2-AcGFP1 vectors containing Il15 (negative control) or 36 distinct murine chemokines and cultured for 18 hours. BFA was added for the final 14 hours to limit chemokine secretion, and cells were analyzed for expression of corresponding chemokine proteins by FC as detailed in Methods. All histograms are gated on GFP⁺ HEK cells comparing Il15 transfectants (gray solid) and respective chemokine transfectants (black tracing) stained with the same chemokine-specific pAb. To reduce proteasomal degradation, Cxcl14-transfected HEK cells were cultured in the presence of 10 μM of the protease inhibitor MG-132 and stained with anti-hCXCL14 mAb clone 131120 or with an anti-hCXCL14 pAb (inset).