Blood -- TAT-mediated intracellular delivery of NPM-derived peptide induces apoptosis in leukemic cells and suppresses leukemogenesis in mice

Blood, Vol. 112, Issue 6, 2474-2483, September 15, 2008

TAT-mediated intracellular delivery of NPM-derived peptide induces apoptosis in leukemic cells and suppresses leukemogenesis in mice
Blood Zhou et al. 112: 2474

Supplemental materials for: Zhou et al

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Figure S1. TAT-NPMΔC suppresses proliferation of Fancc^−/− pre-leukemic stem cells (JPG, 65 KB) -
(A) Clonogenic assay. Fancc^−/− pre-leukemic cells cultured in semi-solid medium containing 30 µg/ml of the indicated proteins were analyzed for serial replating efficiency of hematopoietic progenitors. Data represents the mean ± SD of three experiments. (B) Bone marrow transplantation assay. 1 × 10⁶Fancc^−/− pre-leukemic cells (CD45.2⁺) were transplanted, along with 1 × 10⁶ competitor cells from B6.BoyJ mice (CD45.1⁺), into lethally irradiated recipient (B6.BoyJ) mice, which after 10 days were injected i.p. with the indicated proteins (10 mg/kg in 0.5 ml of PBS and 10% glycerol) twice a week for 4 weeks. Engraftment was evaluated at 4 and 12 weeks post-transplantation. Data represent mean ± SD of three independent experiments, each with 6 animals (total 18 mice).
Figure S2. Retention kinetics of the TAT-fusions in the recipient mice (JPG, 46.3 KB) -
Recipient mice treated with the indicated proteins (10 mg/kg in 0.5 ml of PBS and 10% glycerol) were sacrificed at the indicated time points after the injection, and BM cells were prepared for Western blot analysis using the anti-His antibody. 100 µg of total proteins was loaded in each lane, and equal loading was ensured by probing the blot with anti–β-actin.