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. Author manuscript; available in PMC: 2009 Jun 15.
Published in final edited form as: Free Radic Biol Med. 2006 Mar 30;41(1):86–91. doi: 10.1016/j.freeradbiomed.2006.03.010

Fig. 5.

Fig. 5

H2O2 produced by the ADP-stimulated respiratory burst does not cause glutathionylation of SHP-1 and SHP-2 in NR8383 cells. (A) NR8383 cells were exposed to 400 μM ADP for various times. SHP-1 and SHP-2 were immunoprecipitated with their respective antibodies, and the immunoprecipitates were separated by electrophoresis and immunoblotted with the antibody to glutathione-protein complexes. The membranes were stripped and reprobed with the antibodies to SHP-1 or SHP-2. (B) Disulfide bond formation between SHP-1 and SHP-2 and glutathione was induced by adding 25 mM GSSG in the lysis buffer. Aliquots of lysate were immunoprecipitated with antibodies to SHP-1 or SHP-2. In some samples, DTT was added to the loading buffer to reverse glutathionylation. The immunoprecipitates were separated by electrophoresis and immunoblotted with the antibody to glutathione-protein complexes. The membranes were stripped and reprobed with the antibodies to SHP-1 or SHP-2.