Fig. 7

CASC2 enhances stability of SOCS1 protein. a RIP followed by real-time PCR assay was used to analyze the interaction between CASC2 and SOCS1 protein. Relative quantification of CASC2 in RNA–protein complexes immunoprecipitated with IgG or SOCS1 antibodies from cell extracts. IgG was taken as a negative control. b Western blot of SOCS1 expression in protein complexes pulled down by full-length CASC2 or its antisense control fragment. c Deletion mapping of SOCS1-binding domain in CASC2. Western blot of SOCS1 in protein samples pulled down by different CASC2 fragments. d Control and CASC2 overexpressing KYSE30 cells were treated with CHX (100 μg/ml) for the indicated time points. The cell lysates were examined by immunoblotting. A plot of normalized amount of SOCS1 protein is shown. e Control and CASC2 knockdown KYSE150 cells were treated with CHX (100 μg/ml) for the indicated time points. The cell lysates were examined by immunoblotting. A plot of normalized amount of SOCS1 protein is shown. f Control and CASC2 overexpressing KYSE30 cell lysates were immunoprecipitated with anti-SOCS1 antibody, and the immunocomplexes were immunoblotted with antibodies against Ub. g Control and CASC2 knockdown KYSE150 cell lysates were immunoprecipitated with anti-SOCS1 antibody, and the immunocomplexes were immunoblotted with antibodies against Ub. Error bars indicate SD. *p < 0.05