Figure 6.
ZDHHC12 affects CLDN3 related ovarian tumorigenesis. (A) Confocal images show subcellular localization of endogenous ZO-1and FLAG-tagged CLDN3 in shCtrl and shZDHHC12 A2780 cells. The y–z axis shows the location of ZO-1 and CLDN3 in basal and apical regions. (B) Quantification of correlated colocalization of CLDN3 and ZO-1, representing by the mean of Manders’ coefficient with threshold (tM1 and tM2) (n = 7 for each group; ∗∗∗P < 0.001). (C) Downregulation of phosphorylated ERK1/2 in shZDHHC12 OVCAR8 cells, compared to shCtrl OVCAR8 cells, was indicated by immunoblotting. (D) The tumorigenesis of shCtrl, shZDHHC12#1 and shZDHHC12#2 groups in vivo animal assay according to time course (37 days). The first time that tumor was found in each group was on day 5, day 33 and day 20. In shCtrl group, 7 tumors formed on day 15 which remained to the end of in vivo assay. In shZDHHC12#1 group, 2 tumors formed on day 37 which remained to the end of in vivo assay. In shZDHHC12#2 group, 4 tumors formed on day 35 which remained to the end of in vivo assay. (E) Tumor growth curves of shCtrl, shZDHHC12#1 and shZDHHC12#2 groups in vivo animal assay. Error bars represent SEM (∗∗P < 0.01). (F) Tumor formed in nude mice at the endpoint of in vivo animal assay (n = 8 for each group). At the endpoint of the in vivo animal assay, the tumorigenesis ratio (the number of tumorigenic individuals/the number of total individuals) of each group: 7/8 in shCtrl group, 2/8 in shZDHHC12#1 group, 4/8 in shZDHHC12#2. The tumors were arranged from left to right by weight. (G) The nude mice were sacrificed, and the tumors were removed and measured in weight. Dot plots represent the weight of tumors (n = 8 for each group in all statistical analysis; n.s. indicates no statistic difference; ∗P < 0.05).