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Thiol modification by NEM in intact cells. COS 7 cells were fixed with methanol, permeabilized with triton X-100. For NEM treated cells, thiols were alkylated by a 30 minute incubation with 40 mM NEM in PBS followed by a 30 minute incubation with 40 mM NEM in DMSO. Free thiols were then labeled with Alexa Fluor 488 C5 maleimide as described in methods. DAPI was used for staining nuclei and shown in the lower panels.
