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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: SPAG5 promotes hepatocellular carcinoma progression by downregulating SCARA5 through modifying β-catenin degradation

Fig. 6

SPAG5 regulates SCARA5 expression through the wnt/β-catenin pathway in HCC cells. a, Western blot and qRT-PCR analyses were used to detect SCARA5 expression in Huh7 cells stably transfected with the shNC or the shβ-catenin plasmid. *p < 0.05. b, Western blot analyses were used to detect SCARA5 expression in Hep3B cells stably transfected with the vector or the p-β-catenin plasmid. *p < 0.05. c, Huh7 cells transfected with shNC and shSPAG5 were subjected to western blot analyses for the indicated proteins. d, the relative luciferase activity levels in cells transfected with TOP-flash and FOP-flash vectors in Huh7 cells transfected with shNC and shSPAG5 are shown. *p < 0.05. e, Hep3B cells transfected with vector and p-SPAG5 were subjected to western blot analyses for the indicated proteins. f, the relative luciferase activity levels in cells transfected with TOP-flash and FOP-flash vectors in Hep3B cells transfected with vector and p-SPAG5 are shown. *p < 0.05. g and h, Huh7 cells were transfected with the indicated plasmid. The quantity of SPAG5, β-catenin and SCARA5 were assessed by western blot analysis. The cancer cells’ proliferation capacities were detected by EdU and CCK8 assays. *p < 0.05. i and j, Hep3B cells were transfected with the indicated plasmid. The quantities of SPAG5, β-catenin and SCARA5 were assessed by western blot analysis. The proliferation capacities of the cancer cells were detected by EdU and CCK8 assays. *p < 0.05. k and l, Hep3B cells transduced with vector and p-SPAG5 plasmid were treated with XAV-939. The quantities of SPAG5, β-catenin and SCARA5 were assessed by western blot analysis. The proliferation capacities of the cancer cells were detected by EdU and CCK8 assays. *p < 0.05. m. cells transduced with SPAG5 shRNA or p-SPAG5 plasmid were treated with 10 μM MG132. Cells were collected at 6 h and immunoblotted with the antibodies indicated. n. Huh7 cells were transfected with SPAG5 shRNA or p-SPAG5 plasmid, and treated with cycloheximide (CHX). Cells were collected at different time points and immunoblotted with the antibodies indicated. o. Lysates from Huh7 cells transduced with SPAG5 shRNA or p-SPAG5 plasmid were immunoprecipitated with the anti-Ub and immunoblotted with the anti-β-catenin. Cells were treated with MG132 for 6 h before collection

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