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. 2016 May 20;7(26):40160–40173. doi: 10.18632/oncotarget.9514

Figure 5. Restitution of CDK4 and cyclin D1 rescued the phenotype produced by Tspan5.

Figure 5

A. Reconstitution of CDK4 in Tspan5-overexpressing (Tspan5) or control (Control) tumour cells by transfecting pCMV-Myc-CDK4 into either AGS or MKN45 cells. Western blotting confirming the downregulation of CDK4 by Tspan5 (Tspan5 versus Control) and the upregulation of CDK4 (Tspan5+CDK4 versus Tspan5+Control) in both Tspan5-overexpressing AGS and MKN45 cell lines. B. CCK-8 proliferation assays showing the increased proliferation of both AGS and MKN45 cells after 48 hours by reconstitution of CDK4 in Tspan5-overexpressing cells (ANOVA test, **P<0.001 or ***P<0.001). C. Reconstitution of cyclin D1 in Tspan5-overexpressing (Tspan5) or control (Control) tumour cells by transfecting pENTER-cyclin D1 into either AGS or MKN45 cells. Western blotting confirming the downregulation of cyclin D1 by Tspan5 (Tspan5 versus Control) and the upregulation of cyclin D1 (Tspan5+CycD1 versus Tspan5+Control) in both Tspan5-overexpressing AGS and MKN45 cell lines. D. CCK-8 proliferation assays showing the increased proliferation of both AGS and MKN45 cells after 48 hours by reconstitution of cyclin D1 in Tspan5-overexpressing cells (ANOVA test, **P<0.001 or ***P<0.001).