Generation and characterization of CEP120 constructs

The GFP- or myc-tagged cDNA constructs for CEP120, including those encoding the full-length protein and various CEP120 fragments were as described in our previous paper^{17 (link)}. The GST-fusion constructs were generated by inserting cDNAs encoding various potions of CEP120 in-frame with GST in the pGEX4T vector (GE Healthcare) or were as previously described^{17 (link)}. The QuikChange site-directed mutagenesis kit (Stratagene) was used to make the disease-associated CEP120 mutant constructs. The constructs that express various GFP-CEP120 mutant proteins were generated using pLVX-Tight-puro vectors (BD Biosciences Clontech). The human C2CD3 cDNA fragment, which was a gift from Dr. Gonczy’s lab^{54 (link)}, was subcloned into the pEGFP-C1 vector (BD Biosciences Clontech). The cDNAs encoding full-length Talpid3 or C2CD3 were obtained by RT-PCR from the total RNA of human HEK293T cells, and subcloned in-frame into the pEGFP-C1 vector (BD Biosciences Clontech). The sequences of all constructed plasmids were confirmed.