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Frontiers of Medicine >> 2008, Volume 2, Issue 1 doi: 10.1007/s11684-008-0007-0

Effects of podocin transfection on CD2AP distribution in HEK293 cells

1.Department of Nephrology, Affiliated Nanjing Children's Hospital of Nanjing Medical University; 2.Institute of Pediatrics, Nanjing Medical University

Available online: 2008-03-05

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Abstract

The aim of this paper is to construct a podocin fluorescence expression vector and observe the effects of podocin transfection on CD2AP distribution in HEK293 cells. The pGEMT-easy vector containing the full-length cDNA encoding human podocin was cloned and digested with HI and I. The digested full-length podocin was subcloned into pEGFP-C2. The constructed plasmids were transfected into HEK293 cells and its effects on CD2AP distribution were observed by immunofluorescence. The pEGFP-NPHS2 expression vector was successfully constructed and podocin exclusively located on the HEK293 cell membrane. After podocin transfection, CD2AP redistributed from the perinucleus to the cytoplasm in HEK293 cells. It can be concluded that podocin can recruit CD2AP to redistribute from the perinucleus to the cytoplasm in HEK293 cells.

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