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. 2012 Dec 18;41(2):842–854. doi: 10.1093/nar/gks1255

Figure 2.

Figure 2.

Thr416 (TP5) of EZH2 is the key NIPP1-binding site. (A) Left: EGFP-EZH2 was mutated at the indicated TP-motifs (Thr to Ala mutations) and used for GST pull-down assays with the NIPP1-FHA domain. Right: Quantification and statistical analysis of the immunoblots. Data are shown as means ± SEM (n = 4). Significant differences compared with WT are indicated with *(P < 0.05) or **(P < 0.01) (paired Student’s t-test). (B) Same as in (A) with combined Thr to Ala mutation of the indicated TP-motifs. (C) GST pull-downs of EGFP-EZH2 were performed in the presence of 1 mM of the indicated EZH2 dodecapeptides. (D) Endogenous NIPP1 was immunoprecipitated from HEK293T cell lysates that were transfected with expression vectors for EGFP-tagged EZH2-WT or EZH2-mTP5. The co-immunoprecipitation of the EZH2 fusions was analyzed by immunoblotting with EGFP-antibodies.