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. 2014 Feb 10;15(4):419–427. doi: 10.1002/embr.201338241

Figure 5.

Figure 5

FBI-1 impairs SAM68 binding to BCL-X through histone deacetylase (HDAC) activity.
  1. Cross-link immunoprecipitation (CLIP) analysis of endogenous SAM68 and recombinant Myc-SAM68 in cells transfected or not with Flag-FBI-1. Associated BCL-X RNA was quantified by qRT-PCR and represented as fold enrichment relative to the IgG sample. The scheme indicates the position of primers used for the analysis.
  2. Western blot analysis of HDAC1 co-immunoprecipitated with Flag-FBI-1 or control immunoprecipitates (rabbit IgGs) from HEK293T nuclear extracts.
  3. RT–PCR analysis of endogenous BCL-X mRNA splicing in HEK293T cells treated for 40 h with 50, 100, and 200 nM of LBH589 or with DMSO.
  4. RT–PCR analysis of the BCL-X minigene splicing in HEK293T transfected with or without Flag-FBI-1 and treated for 40 h with LBH or DMSO.
  5. CLIP assay of endogenous SAM68 in HEK293T transfected as in (D). Associated BCL-X RNA was quantified by qRT-PCR and represented as fold enrichment relative to the IgG sample.

Data information: P-values of Student's t-test: *P < 0.05; **P < 0.01; n.s., not significant. Data were obtained from three independent experiments.