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. 2011 Mar 14;30(30):3360–3369. doi: 10.1038/onc.2011.55

Figure 2.

Figure 2

BRC5 repeat of BRCA2 binds directly to HMG20b. (a) BRCA2 and HMG20b complexes were pulled down from 293T extracts using protein A-coupled anti-BRCA2 antibody (αB2-9) and anti-HMG20b antibody (M10), respectively. Coimmunoprecipitated BRCA2 and HMG20b were detected by western blotting. (b) GST–BRCA2 fragments (whose span is shown in the schematic) were incubated with 293T extracts expressing Flag-HMG20b, and the bound proteins were analyzed by western blotting (‘GST pull-down assay'). See also Supplementary Figures S3a and b. (c) The B2-4 fragment was subcloned into five smaller fragments as shown in the schematic, and the GST pull-down assay was performed as shown in (b). Supplementary Figure S3c shows the controls for sample loading. Note that the B2-4 fragment is distinct from the BRC4 repeat. (d) Biotin-labelled BRC4 (1517–1551) and BRC5 (1661–1695) peptides were incubated with 293T extracts expressing Flag-HMG20b (upper panel) and purified HMG20b protein (His-HMG20b, lower panel) in a streptavidin pull-down assay.