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. 2006 Nov 17;103(48):18261–18266. doi: 10.1073/pnas.0606108103

Fig. 4.

Fig. 4.

NOTCH1 binds to c-MYC proximal promoter sequences and regulates c-MYC mRNA and protein expression in T-ALL. (a) Western blot analysis of ICN1 levels in RPMI 8402 T-ALL cells infected with lentiviruses (pLKO puro) driving expression of shRNAs targeting NOTCH1 (shRNA NOTCH1) or control shRNAs targeting the luciferase gene (shRNA LUC). (b) Quantitative RT-PCR analysis of DELTEX1 and c-MYC transcript levels in RPMI 8402 T-ALL cells upon NOTCH1 shRNA knockdown and in control shRNA-treated cells. (c) Western blot analysis of c-MYC in HPB-ALL T-ALL cells showing down-regulation of c-MYC expression upon NOTCH1 inhibition with GSI (CompE 500 nM). The arrowhead indicates the c-MYC-specific band. (d) ChIP analysis of NOTCH1 binding to c-MYC promoter sequences. Quantitative PCR analysis of c-MYC promoter sequences normalized to β-actin levels in control DNA and chromatin immunoprecipitates performed with two NOTCH1 antibodies (N1-TAD and Val 1744) and IgG as control in HPB-ALL cells.