14-3-3 HyA interacts with Hydra Bcl-2-like protein in vitro. (A) bacterially expressed His-tagged 14-3-3 HyA or 14-3-3 HyB were incubated with bacterial lysates containing GST-Hybcl-2-like1 or as a control GST-HyInnexin1 (lanes 1-6). Inputs of 14-3-3 HyA (lane 1) and 14-3-3 HyB (lane 4) represent 12.5% of the total 14-3-3 proteins used in each assay. Eluted 14-3-3 HyA (lanes 2 and 3) and 14-3-3 HyB (lanes 5 and 6) are shown after pulldown with GST-fusion-proteins. Western blots were probed with anti-Xpress antibody, which recognises the His-tag of the bacterially expressed 14-3-3 proteins. In (B) Western blots with samples from (A) were probed with anti-GST antibody to show that equal amounts of GST-fusion-protein were eluted in all assays. Asterisks indicate full-length GST-fusion proteins. Smaller fragments represent degradation products.
Pauly et al. BMC Cell Biology 2007 8:31 doi:10.1186/1471-2121-8-31