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. 2021 Mar 31;11(12):5759–5777. doi: 10.7150/thno.57659

Figure 3.

Figure 3

HOXB5 promotes HCC metastasis through transactivating CXCL1 expression. (A) The diagram showed the genes regulated by HOXB5 changes. (B) RT-qPCR and ELISA were used to detect the CXCL1 expression. (C) Luciferase reporter assay was performed after co-transfection of CXCL1 promoter luciferase construct and pCMV-HOXB5. (D) Cells were first co-transfection of pCMV-HOXB5 and truncated and mutated CXCL1 promoter and relative luciferase activity was measured. (E) ChIP assays revealed the binding of HOXB5 in CXCL1 promoter in HCC cell lines and HCC specimens. (F) Western blotting and ELISA analyzed protein expression of CXCL1 in the indicated HCC cells. (G-K) In vivo assays showed that HOXB5 promoted HCC metastasis through upregulating CXCL1. (G-H) The C57BL/6 mice were implanted with the indicated cells in the liver. Bioluminescent images and growth rate were shown. (I) Overall survival time of C56BL/6 mice in different groups was shown. (J) Lung metastatic nodules were counted. (K) Lung metastatic nodules in different groups. * P < 0.05.