Fig. 2

Increased SIRT1 protein levels in T-ALL with Notch1 mutation. a-d Immunoblots and relative quantification expression of SIRT1 in MOLT-4 and CCRF-CEM cells treated with GSI, then washed and refed medium containing DAPT (mock) or medium lacking DAPT (wash). e-f Western blotting for SIRT1 and MYC levels in MYC-ShRNA-expressing MOLT-4 and CCRF-CEM cells. g-h Western blotting for SIRT1 following CHX treatment of MYC knockdown or control MOLT-4 and CCRF-CEM cells (left panel). The right panel showed results of densitometry quantitation (n = 3). i 293 T cells were co-transfected with pCMV-N-Flag (Flag-Vector, FV), Flag-SIRT1, pCMV-Blank (Vector, V), pCMV-MYC and HA-tagged ubiquitin as indicated and subjected to ubiquitination analysis