MOESM5 of Citrullination of HP1γ chromodomain affects association with chromatin

Additional file 5: Figure S4. Differentiation of mESCs. A Immunoprecipitation (IP) of endogenous HP1γ from nuclear lysates of mESCs before and after 72 h LIF withdrawal. IPs were performed with anti-HP1γ and anti-HA control antibodies and analysed by immunoblotting using an anti-peptidyl-citrulline antibody (α-Citrulline). Subsequently the same immunoblots were stripped and re-probed with an anti-HP1γ antibody (α-HP1γ). 4% of input amounts of each IP are indicated. Replicate (i) is shown in Fig. 4D. B Stable exogenous expression of mEos3.2–HaloTag–HP1γ fusion proteins does not affect total mRNA level of pluripotency markers in mESCs. RT-PCR data for the indicated genes were normalised to β-actin mRNA expression. Bars represent ± SEM (n = 2). C The mRNA levels of pluripotency markers in mESCs decrease after withdrawal of LIF. The mRNA levels of the indicated genes were measured by RT-PCR over a course of 3 days after withdrawal of LIF. RT-PCR data were normalised to β-actin mRNA expression, and expression fold change was determined relative to d0 time point using the ddCT method. Bars represent ± SEM (n = 2). D Representative light field microscope images of mESCs before and after 72 h LIF withdrawal, captured with a Leica EC3 camera at 20 × magnification. Scale bars: 100 μ