Figure 1.

SSH-1L participates in H₂O₂-induced cofilin dephosphorylation. (A) HeLa cells were pretreated with or without 0.1 mM Na₃VO₄ for 30 min and then stimulated with 0.5 mM H₂O₂ for the indicated times. Cell lysates were immunoblotted with antibodies to phosphorylated cofilin (p-Cofilin) or cofilin (t-Cofilin). (B) HeLa cells were pretreated with or without the indicated inhibitor sets (1 mM Na₃VO₄, 1 μM okadaic acid [OA], 1 μM CalyculinA [CalA], 30 μM BAMTA-AM, 1 mM EGTA) for 0.5 h and then stimulated with 0.5 mM H₂O₂ for 30 min. Cell lysates were immunoblotted with antibodies to phosphorylated cofilin (p-Cofilin) or cofilin (t-Cofilin). (C) HeLa cells were transfected with 10 nM control, CIN or SSH-1L siRNA (si) for 48 h and then stimulated with or without 0.5 mM H₂O₂ for 30 min. Cell lysates were immunoblotted with antibodies to phosphorylated LIMK1/2 (p-LIMK), phosphorylated cofilin (p-Cofilin), cofilin (t-Cofilin), SSH-1L, CIN, or actin. (A–C) The graphs at the right of each panel represent the averaged normalized p-cofilin values; the value in Na₃VO₄ + H₂O₂-treated cells is taken as 1.0. Values are from three independent experiments ± SD.