Figure 6.

(A) Pull down assays were performed with GST alone or GST fused to full length Rb (GST-RB) as described previously.⁴⁸ MDA-MB-231 cell lysate (500µg) was incubated with GST or GST-RB for 2 hours and associated proteins were collected using GST-sepharose. Immunoblotting with ZEB1 antibodies is shown. Data shown is representative of 3 separate experiments. (B) MDA-MB-231 cells were grown in 2D cell culture and subjected to PNUTS depletion. Co-immunoprecipitation was performed using ZEB1 antibodies using cells subjected to NT (nontargeting) siRNA or PNUTS siRNA followed by immunoblotting with antibodies to PNUTS, ZEB1 and Actin. Phospho-Rb and unphospho-Rb are separated by differential migration on SDS-PAGE and are indicated. Data shown is representative of 2 independent experiments.