The interaction of RACK1 with CLIC1 in cells / 安徽医科大学学报

ABSTRACT

Objective @#To investigate the interaction between intracellular chloride ion protein 1(CLIC1) and activated protein kinase C receptor 1 ( RACK1) .@*Methods @#Plasmids pcDNA3. 1⁃RACK1⁃HA and/or pcDNA3. 1 ⁃CLIC1⁃FLAG were transfected into HEK 293T cells, and pcDNA3. 1⁃RACK1⁃HA and/or pcDNA3. 1⁃CLIC1⁃FLAG were transfected into COS7. GST⁃pulldown and immunoprecipitation assays were performed to determine the interaction between CLIC1 and RACK1 in vivo and in vitro. The co⁃localization of CLIC1 and RACK1 was observed by indirect immunofluorescence assay.@*Results @#Western blot confirmed that CLIC1 and RACK1 could be highly expressed in HEK 293T cells. GST⁃pulldown showed that RACK1 bound CLIC1 in vitro, and immunoprecipitation showed that CLIC1 and RACK1 interacted in vivo. Furthermore, indirect immunofluorescence assay showed CLIC1 co⁃localized with RACK1 .@*Conclusion @#Human CLIC1 protein interacts with RACK1 in vitro and in vivo.