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. 2009 Jul 9;106(30):12301–12305. doi: 10.1073/pnas.0901823106

Fig. 4.

Fig. 4.

PGAM5 dephosphorylates ASK1. (A) PGAM5 induces a mobility shift of the band of ASK1. Deletion mutants of ASK1 [CT, amino acids (aa) 956–1374; ΔC, amino acid 1–940; ΔN, amino acid 649–1374] with or without Flag-PGAM5 were expressed in HEK293 cells. Cell lysates were subjected to immunoblotting (IB). Schematic illustrations of ASK1 and its deletion mutants are also shown. (B) ASK1-CT possesses phosphorylation site(s) targeted by PGAM5. Cell lysates from HEK293 cells expressing HA-ASK1-CT with or without PGAM5 were left untreated or treated with λPPase and subjected to IB. (C) PGAM5 dephosphorylates ASK1-CT in vitro. Flag-ASK1-CT was expressed in HEK293 cells and immunoprecipitated with anti-Flag antibody. After incubation with GST-PGAM5-WT, -H/A, -H/F, or λPPase, the immunoprecipitates were subjected to IB.