Figure 6.

Ik1 and Ik6 display distinct effects on chromatin histone deacetylation on the FGFR4 promoter. Chromatin immunoprecipitation (ChIP) was performed as described in Materials and Methods. a: DNA from lysate before immunoprecipitation (input DNA) or from plasmid FGFR4 (P−115/+99-Luc) DNA were amplified as positive controls. GH4 cells were co-transfected with 5′ FGFR4 promoter in combination with expression vectors encoding either Ik1, Ik6, or empty vector control. Cross-linked chromatin was immunoprecipitated with antibody to acetyl histone 3 (AcH3) and subjected to PCR analysis. Note the effect of Ik1 on histone deacetylation resulting in absence of PCR product from AcH3 antibody immunoprecipitates. Omission of AcH3 antibody immunoprecipitation (non-IP) results in absence of PCR products from all corresponding transfections. b: Corresponding immunoblotting with AcH3 antibody of immunoprecipitates reveals no effect or difference between Ik1 and Ik6 on global histone acetylation in pituitary cells.