Figure 5
CEP120 interacts with C2CD3 and Talpid3. (a) CEP120 physically associates with C2CD3 in vivo. HEK293T cells were co-transfected with CEP120-myc and GFP-C2CD3 constructs. Twenty-four hours after transfection, cell lysates were immunoprecipitated (IP) and then immunoblotted (IB) with the indicated antibodies. (b) Mapping the C2CD3-interacting domain in CEP120. HEK293T cells were co-transfected with GFP-C2CD3 and various myc-tagged CEP120 constructs, analyzed by IP, and subsequent IB using the indicated antibodies. (c) CEP120 directly interacts with C2CD3. GST and various GST-CEP120 recombinant proteins were affinity purified (left). The full-length 35S-methionine-labeled C2CD3 proteins (middle) were incubated with bead-bound GST or various GST-CEP120 recombinant proteins (right) and analyzed by SDS-PAGE and autoradiography. (d) CEP120 physically associates with Talpid3 in vivo. HEK293T cells were co-transfected with CEP120-myc and GFP-Talpid3 constructs, and analyzed as described in (a). (e) Mapping the Talpid3-interacting domain in CEP120. HEK293T cells were co-transfected with GFP-Talpid3 and various myc-tagged CEP120 constructs and analyzed as described in (b). Uncropped blots of (a,b,d,e) are shown in Fig. S6b–e, respectively. (f) CEP120 directly interacts with Talpid3. The full-length 35S-methionine-labeled Talpid3 proteins (left) were incubated with bead-bound GST or various GST-CEP120 recombinant proteins (right) and analyzed by SDS-PAGE and autoradiography. (g) Schematic summary of the interactions of CEP120, C2CD3, and Talpid3, and the positions of the disease-associated CEP120 mutations that cause JS.
