Fig. 8.

In-gel profiling reveals acylated proteins in RAW264 cells. (A) Scheme of experimental procedure comprising preincubation of cells with 150 μm or 500 μm palmitic acid (30 min, 37 °C; PA) or 125 μm or 250 μm BPA (1 h, 37 °C) or with 30 μm BSA in controls followed by metabolic labeling of cells with 50 μm 17ODYA or exposed to 0.05% DMSO carrier (- 17ODYA) for 4 h. After lysis, click chemistry reaction was performed with IRDye 800CW-azide and proteins were separated by SDS-PAGE. A set of samples was exposed to 2.5% hydroxylamine (HXA, 30 min, 25 °C) before SDS-PAGE. (B, C) Profile of 17ODYA-labeled proteins in cells exposed to PA (B) or BPA (C) revealed by in-gel fluorescence (left panels) and stained with Coomassie Blue to verify equal loading (right panels). Molecular weight markers are shown on the right.