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. 2011 Sep 29;6(9):e25358. doi: 10.1371/journal.pone.0025358

Figure 1. SOCS2 forms a stable complex with Elongin B and C, Cullin 5 and Rbx2 mediated by the SOCS-box.

Figure 1

(A) Schematic of the various SOCS-constructs that were used. (B) Hek293T cells were transfected with Elongin B, Elongin C and Myc-Cullin5 and either an empty vector (lane 1), wildtype (lane 2 and 8) or a mutant form of FLAG-SOCS2 (lanes 3–7) as denoted in the picture. FLAG-SOCS2 and FLAG-SOCS2 mutants were immunoprecipitated with FLAG-beads (lanes 1–7), Mock-ip was immunoprecipitated with IgG. Western blots of immunoprecipitates and the lysate was performed as described in the Materials and Methods. (C) and (D) Western blot of whole cell lysate from HEK293T cells transfected with FLAG-SOCS2 and both, either or no Elongins as detailed in the figure. (E) HEK293T cells transfected with either FLAG-SOCS2 and both Elongins or FLAG-SOCS2 alone were treated with cycloheximide (100 µg/ml) for the times indicated in the figure before lysis. C denotes the control which was transfected with Empty vector instead of SOCS2.