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. 2018 May 8;11:2581–2591. doi: 10.2147/OTT.S162387

Figure 2.

Figure 2

WWC3 level correlates with the expression of EMT process biomarkers.

Notes: (A and C) H1299 cells were transfected with WT WWC3 or mutant plasmid (−ΔWW) and an EV, and 48 h later, cells were lysed and the proteins and mRNA were detected by immunoblotting and RT-qPCR, respectively. GAPDH expression served as a loading control. (B and D) Knockdown of WWC3 in A549 cells using two independent shRNAs had the opposite effect. (E) Silencing WWC3 enhanced N-cadherin expression. A549 cells were transfected with shRNA-WWC3 or shCTL for 48 h and then analyzed for endogenous N-cadherin with confocal microscopy (magnification, 400×). (F) The representative photographs of the expression correlation between WWC3 and E-cadherin and N-cadherin in human lung cancer specimens (magnification, 400×). All experiments were performed in triplicate. Data represent the mean±SD. *P<0.05.

Abbreviations: WWC3, WW and C2 domain containing protein-3; EMT, epithelial–mesenchymal transition; WT, wild type; EV, empty vector; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; shCTL, short-hairpin RNA control; ns, nonsignificant; DAPI, 4′,6-diamidino-2-phenylindole.