Fig. 5

Expression of FBXW10 is regulated by ZNF169, and FBXW10 knockdown inhibits cell proliferation. (A) Correlation between ZNF169 and FBXW10 expression (n = 510). THCA, thyroid carcinoma. (B) RT-qPCR analyses of FBXW10 expression in K1 cells transfected with the ZNF169 overexpression plasmid and Ctrl, and BCPAP and TPC1 cells transfected with shCtrl, shZNF169#1 and shZNF169#2. β-actin was used for normalization. (C) Western blot analyses of FBXW10 expression in K1 cells transfected with the ZNF169 overexpression plasmid and Ctrl. (D) Western blot analyses of FBXW10 expression in BCPAP and TPC1 cells transfected with shCtrl, shFBXW10#1 and shFBXW10#2. GAPDH was used as the loading control. (E) The binding relationship between ZNF169 and the FBXW10 promoter was analyzed by dual-luciferase reporter assay. (F) qPCR analysis shows FBXW10 levels that are pulled down in ChIP assay with the anti- ZNF169 antibody or IgG. The qPCR product was subjected to agarose gel electrophoresis. K1 cell lysates were used for the ChIP assay. (G) The knockdown efficiency of FBXW10 in TPC1 cells transfected with shCtrl, shFBXW10#1 and shFBXW10#2. (H) Cell Counting Kit-8 analysis of TPC1 cells transfected with shCtrl, shFBXW10#1 and shFBXW10#2. (I) Colony formation assay of TPC1 cells transfected with shCtrl, shFBXW10#1 and shFBXW10#2. (J) The knockdown efficiency of FBXW10 in BCPAP cells transfected with shCtrl, shFBXW10#1 and shFBXW10#2. (K) Cell Counting Kit-8 analysis of BCPAP cells transfected with shCtrl, shFBXW10#1 and shFBXW10#2. (L) Colony formation assay of BCPAP cells transfected with shCtrl, shFBXW10#1 and shFBXW10#2. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001