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Figure 4


Fig. 4. Stability of the CIS and SOCS2 interactions with the LR. (A) HEK293T cells were transiently co-transfected with plasmids encoding different pMET7-LR variants, the pMG2-CIS or pMG2-SOCS2 prey construct, pEF-FLAG-I/mCIS or pEF-FLAG-I/mSOCS2, or the appropriate amount of mock vector together with the pXP2d2-rPAP1-luci. The transfected cells were either stimulated for 24 hours with leptin or were left untreated (NS, not stimulated). Luciferase measurements were performed in triplicate. Data are expressed as mean fold induction (leptin stimulated/NS) + s.d. (B) Western blot analysis of CISprey, SOCS2prey, CIS and SOCS2 expression. Expression of the FLAG-tagged fusion proteins, CIS and SOCS2 was verified on lysates of transfected cells using anti-FLAG antibody.