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. Author manuscript; available in PMC: 2019 Mar 12.
Published in final edited form as: Biochem Biophys Res Commun. 2013 Sep 25;440(3):431–436. doi: 10.1016/j.bbrc.2013.09.090

Fig. 1.

Fig. 1

Fem1b interacts with Gli1. (A) Gli1 co-immunoprecipitates with Fem1b. HA-Fem1b and Myc-Gli1 or empty Myc vector were co-transfected into 293T cells and immunoprecipitation was performed after 24 h with a Myc antibody or control IgG followed by Western blot. Input represents 8% of the whole cell lysate (WCL). (B) Recombinant Fem1b binds ectopically expressed Gli1. GST pull-down was performed with GST or GST-Fem1b using 293T lysate expressing Myc-Gli1 20 h post-transfection followed by Western blot. Ponceau-S demonstrates the relative amount of GST fusion bait protein. Input represents 0.25% of total protein. (C) Recombinant Fem1b binds endogenous Gli1. GST pull-down was performed with the indicated constructs using RMS-13 lysate followed by Western blot. Input represents 0.25% of the total protein and (D) Fem1b and Gli1 interact directly. GST pull-down was performed with the indicated constructs using human Gli1 programmed rabbit reticulocyte lysate. Input represents 1.5% of the total rabbit reticulocyte protein.