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Fig. 6 | Biology Direct

Fig. 6

From: A functional reference map of the RNF8 interactome in cancer

Fig. 6

In vitro validation of the interaction between RNF8 and YBX1. A The intersection of RNF8 interactome by LC–MS and potential RNF8-interacting proteins acquired using five online tools. B Schematic diagram of Co-IP. C HEK-293T cells were transfected with pcDNA3.1 (control) or pcDNA3.1-FH-RNF8 and harvested after 48 h of transfection. HEK-293T cell lysates were incubated with HA-Sepharose beads, and the bound proteins were analyzed via western bolting with anti-RNF8, anti-HA, and anti-YBX1 antibodies. D we harvested MDA-MB-231 cells, then the lysates were co-immunoprecipitated with anti-RNF8 antibody or normal mouse IgG(control), and the elution protein complexes were analyzed by western blotting with anti-RNF8 and anti-YBX1 antibodies. E HEK-293T cells were transfected with TG006 or TG006-RNF8 with pcDNA3.1-FH-YBX1. At 48 h post-transfection, the cells were harvested and co-immunoprecipitated with anti-Flag-M2 agarose beads, and the YBX1 protein complexes were analyzed by western blotting with anti-YBX1 and anti-RNF8 antibodies. F HEK-293T cells were serially transfected with the corresponding plasmid. At 72 h post-transfection, the cells were harvested and coimmunoprecipitated with anti-Flag-M2 agarose beads, and the YBX1 protein complexes were analyzed by western blotting with anti-Myc (Ub), anti-YBX1, and anti-RNF8 antibodies

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