Expression And Localization Of Myopodin Protein In Cells And Tissues

Myopodin is the second member of the synaptopodin protein family. It shows no significant homology to any known protein except synaptopodin. myopodin contains two classic nuclear localization signals (NLSs), at its N-terminus and C-terminus respectively.There is an actin-binding site at the position of aa 410-563 that can interact with actin directly. Myopodin is a dual compartment protein which redistributes in both the nucleus and the cytoplasm in a differentiation-dependent and stress-induced fashion. To study the function of Myopodin protein, we develop a anti-Myopodin antibody. A recombinant protein containing 1077bp N-terminal sequence of N-Myopodin fused to a GST tag was expressed in E.coli. By immunizing rabbits with the purified recombinant protein, a rabbit anti-N-Myopodin antibody was obtained.Western blots analysis indicated the specifity for this antibody.the distribution of myopodin was analyzed by immunofluorescence microscopy in HeLa,T24, MG63, NIH3T3 and 293T cells using polyclonal anti-N-Myopodin antibody. Myopodin was found in both the nucleus and cytoplasm of MG63 cells, while only appear in the cytoplasm of T24 and HeLa cells, and strictly distribute in the nucleus of NIH3T3 and 293T cells. By immunohistochemistry, Myopodin was found widely expressed in tissues including gullet, stomach and tharm, where its distribution was found in the cytoplasm of both normal and cancerous cells. It was the first time that the distribution and expression of Myopodin was observed across different cell lines. By immunohistochemistry and Western blot, we found Myopodin is expressed in the cytoplasm of both normal and cancerous liver cells. However, its expression significantly decreased in cancerous Liver...