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. 2007 Mar 22;26(8):2083–2093. doi: 10.1038/sj.emboj.7601652

Figure 3.

Figure 3

Substitution of Ser612 with Ala decreases the binding of pRB to E2F-1 and the transcriptional repression. (A) Substitution of Ser612 with Ala decreases pRB–E2F-1 binding. SAOS2 cells were transiently transfected with the indicated constructs. After 40 h, cell lysates were immunoprecipitated with anti-Flag antibody and immunoblotted with anti-myc antibody (top) or anti-Flag antibody (middle). The expression level of pRB was assessed by the immunoblotting of immunoprecipitates with anti-pRB antibody (bottom). (B) Transcriptional repressive activity of pRB S612A is lower than that of wild-type pRB. MCF7 cells were co-transfected with the p73-Luc reporter gene, E2F-1, DP-1, and pGL4.74[hRluc/TK] in the absence or presence of pRB. After 24 h, the luciferase activity in cell lysates was measured and was normalized with the Renilla luciferase activity. The change in the mean±s.d. based on three cultures is shown.