Fig. 2.

Experimental design and quality control of whole-cell (WC) and affinity purification (AP) samples prior to mass spectrometry (MS).A, schematic of experimental design. Transduced and untransduced BV2 and N2A cells were treated with biotin and lipopolysaccharide (LPS) or PBS for 48 h. WC lysates and streptavidin-affinity purified (AP) samples were processed in parallel with MS. B, Western blot (WB) and Coomassie confirming biotinylation of proteins in transduced BV2 WC lysates. C, WB and Coomassie confirming biotinylation of proteins in transduced N2A WC lysates. D, WB and silver stain confirming biotinylation of proteins bound to streptavidin beads and specificity of streptavidin beads for biotinylated species in BV2 AP preparations. LPS impacts banding patterns (10–40 kDa) in transduced BV2 biotinylated proteins visualized with both silver stain and WB. E, WB and silver stain confirming biotinylation of proteins bound to streptavidin beads and specificity of streptavidin beads for biotinylated species in N2A AP preparations.