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. 2014 Dec 10;5(22):11695–11708. doi: 10.18632/oncotarget.2586

Figure 5. GKN1 directly binds to c-myc and inhibits c-myc-induced hTERT expression.

Figure 5

(A) GKN1 stable transfectants from passage 5 were transiently transfected with c-myc. GKN1 completely inhibited the binding activity of c-myc to a hTERT gene promoter in the ChIP assay. (B) Quantitative ChIP-qPCR assay of hTERT revealed that hTERT promoter occupancy increased in c-myc-transfected cells. However, it was strongly reduced in GKN1 stably expressed cells. (n = 3; error bars represent SEM; *p < 0.05 for a one-sided paired t-test). (C and D) Immunoprecipitated c-myc or TRF1 proteins were resolved by SDS-PAGE, transferred to PVDF membranes, and processed for immunoblotting with the indicated antibodies. The GKN1 protein in stable AGSGKN1 cells bound to c-myc and reduced its expression. In addition, the TRF1 protein bound to the c-myc protein in stable AGSGKN1 cells transiently transfected with c-myc and TRF1.