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Figure 4 | BMC Biochemistry

Figure 4

From: p42MAPK-mediated phosphorylation of xEIAP/XLX in Xenopus cytostatic factor-arrested egg extracts

Figure 4

Phosphorylation of Ser235/251/254 affects neither protein stability nor apoptosis-inhibiting activity of xEIAP/XLX. (A) Stability and electrophoretic mobility shift of xEIAP/XLX. The WT/3A/3D forms of 35S-radiolabeled 6XHis-FL, MBP-FL, and MBP-Δ1 were incubated in CSF-arrested (CSF) or interphase (INT) egg extracts for indicated time periods (0, 2, and 4 h). After separation by SDS-PAGE, radiolabeled and MBP-fused recombinants were detected by image analyzer and Western blot, respectively. Molecular weights of standard proteins are indicated at the left. The larger (~70 k) and smaller cleavage products (50–55 k) of MBP-fusions are indicated by single and double asterisks, respectively. (B) Apoptosis inhibition by MBP-FL. (C) Apoptosis inhibition by MBP-Δ1. In (B) and (C), nuclear fragmentation assays were carried out as previously described [11]. Open circles, MBP (negative control); closed circles, WT; open triangles, 3A; closed triangles, 3D; open squares, MBP-xXIAP (positive control). Data are represented as mean + S.E.M. (N = 5).

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