Fig. 3
a H4.WT cells and H4.MetRSL274G cells were transiently transfected with αsyn-RFP or αsyn-eGFP, respectively. Media were supplemented with 4 mM ANL for 72 h. Conditioned media from each cell type was concentrated, and subjected to our BONCAT and affinity purification pipeline, separately, as well as after mixing in equal proportions to simulate co-culture. b Silver stain of eluates show only minimal background labeling from H4.WT cell conditioned media, abundant protein labeling from H4.MetRSL274G cell conditioned media, and intermediate labeling from the simulated co-culture conditioned media. c Immunoblotting unprocessed concentrated conditioned media and post-BONCAT eluates for RFP and GFP shows our pipeline results in retention of GFP from the MetRS expressing cells, but not RFP from non-MetRSL274G-expressing cells. This demonstrates that our pipeline can be applied to secreted proteins in conditioned media. Uncropped blots are available in Additional file 3: Fig. S3