Fig. 5.

Protein Adduction inDhcr7-deficient Neuro2a Cells. A. Workflow to identify lipid-adducted proteins. Intensity oown in A. C. Enrichment Ff Streptavidin indicates the degree of protein adduction (A1). Photo-eluted proteins were visualized in short stacking SDS Page gel shown in A2 followed by in-gel trypsin digestion for LC-MS/MS analysis. 10 and 5 μM of a-Lan and a-7-DHC were supplemented respectively. B. Number of total proteins identified from experiments shactor for the 423 common proteins adducted by both a-DHC and a-Lan. Enrichment factor for a-7-DHC (in black) vs. treatment with DMSO and a-Lan (in blue) vs. treatment with DMSO. The enrichment factor is defined as log₂ of [spectral count for a protein found for alkyne treatment divided by spectral count of the same protein for DMSO treatment]. Four proteins involved in cholesterol biosynthesis were identified in both the a-7-DHC and a-Lan experiments and are highlighted in red in the Figure. Additional replica data of Streptavidin western and stacking gels are included in Supporting Information (Fig. S2). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)