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. 2013 Nov 25;104(12):1647–1655. doi: 10.1111/cas.12303

Figure 2.

Figure 2

Ubiquitination of Hbo1 and promotion of estrogen receptor α (ERα) ubiquitination by Hbo1. (a) Hbo1 (lanes 2, 4 and 6) and the empty vector (lanes 1, 3 and 5), together with HA‐tagged ubiquitin, were transfected into 293T cells. Two days post‐transfection, the cells were lysed under denaturing conditions and immunoprecipitated with anti‐Hbo1 antibody. The immunoprecipitates were assayed for Hbo1 (lanes 3 and 4) and HA peptide (lanes 5 and 6). Ten percent of the input was analyzed. *Immunoglobulin heavy chain. (b) Components of the Hbo1 protein complex. (c) 293T cells were transfected with ERα, Hbo1, Jade‐1, ING4 or ING5, with HA‐tagged ubiquitin by the listed combination. Immunoprecipitation with anti‐ERα antibody was done similarly to Figure 2(a). The immunoprecipitates were probed with anti‐ERα and anti‐HA antibodies. Ten percent of the input was analyzed. *Immunoglobulin heavy chain.