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Fig. 4 | Cancer Cell International

Fig. 4

From: FBXO2 modulates STAT3 signaling to regulate proliferation and tumorigenicity of osteosarcoma cells

Fig. 4

FBXO2 interacts with interleukin 6 receptor to stabilize it. a The interaction protein network of FBXO2 revealed by the BioGRID database. b 293T cell were transfected with EV, Flag-FBXO2 WT or MUT for 36 h. Cells were lysed and immunoprecipitated with Flag M2 beads. The immunoprecipitates were detected by western blot using indicated antibodies. c Beads coated with bacterially expressed GST or GST-SENP2 were incubated with purified Flag-IL6ST or Flag-IL6Ra protein. Beads were washed, and the bound proteins were analyzed by Western blotting with indicated antibodies. d Immunoblotting analysis of the cell lysates of MG63 cells transfected with increased doses of FBXO2WT. e qRT-PCR analysis of IL6ST and FBXO2 in MG63 cells transfected with EV or Flag-FBXO2 WT. f Cell lysates from FBXO2 WT and KO U2OS cells treated with 20 μg/ml cycloheximide (CHX) were subjected to western blot with the indicated antibodies. g U2OS cells were co-transfected STAT3-Luc with indicated plasmids. 24 h after transfection, cells were treated with DMSO or IL-6 (5 ng/ml) for additional 12 h. The cells were lysed and assayed for luciferase assay as above described. **p < 0.01

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