Figure 3.
Association of SETD5 with HDAC3 and PAF1 Complex Components
(A) Silver staining of an SDS-PAGE gel loaded with an immunoprecipitate of 3×FLAG-tagged human SETD5 expressed in HEK293T cells. An immunoprecipitate prepared from cells transfected with the corresponding empty vector served as a control. Proteins identified by LC-MS/MS analysis are indicated.
(B) Silver staining of an SDS-PAGE gel loaded with an immunoprecipitate of 3×FLAG-SETD5 expressed in SH-SY5Y cells with the use of the doxycyline-inducible system. An immunoprecipitate prepared from corresponding cells not treated with doxycycline served as a control. Proteins identified by LC-MS/MS analysis are indicated.
(C) Immunoblot (IB) analysis of the indicated proteins in fractions obtained by gel filtration of lysates of SH-SY5Y cells expressing 3×FLAG-SETD5.
(D) Lysates of HEK293T cells expressing full-length 3×FLAG-SETD5 or the indicated deletion mutants thereof (or transfected with the corresponding empty vector) were subjected to immunoprecipitation (IP) with antibodies to FLAG, and the resulting precipitates as well as the original cell lysates were subjected to immunoblot analysis with antibodies to the indicated proteins.