Research Paper Volume 14, Issue 13 pp 5493—5510

Figure 7. RBX1 promotes the TWIST1-mediated migration dependent on FBXO45. (A, B) The qRT-PCR and western blotting of the TWIST1 expression levels in the SUM159PT cells stably transfected by shFBXO45 or shNC plasmid. *P < 0.05. (C, D) The qRT-PCR and western blotting of the TWIST1 expression levels in the MDA-MB-231 cells stably transfected by vector or plasmid with the overexpression of FBXO45. *P < 0.05. (E) Western blotting was applied for detecting the levels of TWIST1 and FBXO45 protein in TNBC cancer tissues together with paired non-tumour tissues, 30 numbers in each group. Tubulin was utilized as a loading control. (F) Scatter plots of TWIST1 and FBXO45 protein expression in the TNBC. (G) Western blot of TWIST1, FBXO45, and RBX1 protein levels in the MDA-MB-231 cells stably transfected by shRBX1 with or without shFBXO45. The expression of tubulin was employed as a loading control. (H, I) Transwell analyses were implement to identify the MDA-MB-231 cells metastatic capacity that stably transfected by shRBX1 with or without shFBXO45. *P < 0.05. (J) The data reflect the statistics of the number of nodules of lung metastases. *P < 0.05. (K) Western blot of TWIST1, FBXO45, and RBX1 protein levels in the TNBC cells stably transfected by p-RBX1with or without p-FBXO45. The expression of tubulin was employed as a loading control. (L, M) Transwell analyses were implement to identify the TNBC cells metastatic capacity that stably transfected by p-RBX1 with or without p-FBXO45. *P < 0.05. (N) The data display the statistics of the number of nodules of lung metastases. *P < 0.05. (O) Schematic representation of E3 UB ligase RBX1 driving the transfer of TNBC via an FBXO45-TWIST1-dependent degradation mechanism.