cDNA cloning

Human NUP93 full-length cDNA was subcloned by PCR from human full-length cDNA (cDNA clone MGC: 21106 IMAGE: 4750923). Mouse NUP93 was subcloned from mouse NUP93 full-length cDNA (cDNA clone MGC: 28230, IMAGE: 3991335). Human NUP205 subcloned from human full-length cDNA (cDNA clone MGC: 168237 IMAGE: 9020614). Human XPO5 full-length was subcloned from human full-length cDNA (cDNA clone MGC: 74566 IMAGE: 5492046). Human SMAD4 full-length was subcloned from human full-length cDNA (cDNA clone MGC: 8602 IMAGE: 2961238), human SMAD2 was subcloned from human full-length cDNA (cDNA clone MGC: 34440 IMAGE: 5221801), human SMAD5 was subcloned from human full-length cDNA (cDNA clone MGC: 8960 IMAGE:3906006). Mouse Ipo7 was subcloned from mouse full-length Importin7 (cDNA clone, MGC: 175386, IMAGE: 100014508). Expression vectors were produced using LR clonase (Invitrogen®) following the manufacturer's instruction. The following expression vectors were used in this publication: pRK5-N-Myc, pCDNA6.2-N-GFP, pCDNA6.2-C-GFP, pCS-Dest2, and pSirenRetroQ. Clones reflecting the mutations identified in individuals with SRNS were introduced in the cDNA constructed using the Quick change II XL site-directed mutagenesis kit, Agilent Technologies. This technique was also applied to generate a human SMAD4 construct lacking the canonical nuclear export signal (amino acids 142-149).⁴⁵