Shh n peptide

SHH-N peptide (R&D Systems) and SAG (CAS #2095432-58-7, Selleckchem) were used to exogenously induce Shh pathway activity. The potent Smoothened inhibitors cyclopamine (CAS #4449-51-8) and vismodegib (CAS #879085-55-9) were purchased from LC Laboratories. Additional Shh pathway disruptors assessed include U18666A (CAS #3039-71-2, Tocris), RU-SKI 43 (CAS # 1782573-67-4, Tocris), the anti-SHH monoclonal neutralizing 5E1 antibody (Developmental Studies Hybridoma Bank at the University of Iowa), piperonyl butoxide (CAS #51-03-6, Toronto Research Chemicals), and GANT61 (CAS # 500579-04-4, Tocris). The negative control compound benzo[a]pyrene was purchased from Sigma-Aldrich (CAS #50-32-8). All chemicals were dissolved in DMSO or water.

The MUTZ-1 cell line was provided by the Institute of Hematology of Zhejiang University (Hangzhou, China) and was established from the peripheral blood of a 5-year-old girl with MDS French-American-British (FAB) subtype refractory anemia with excess of blasts (RAEB) and progressed quickly to an acute myeloid leukemia (FAB M2). Del (5q) and additional rearrangements involving chromosome 5 [der (15) t (5; 15)] were detected [21 (link)]. Cells were cultured in RPMI-1640 medium using 10% fetal bovine serum and 1% penicillin-streptomycin (Gibco, Life Technologies) at conditions of 37°C, 100% humidity, and 5% CO₂.
Shh-N peptide dissolved in phosphate-buffered saline was purchased from R&D Systems, USA. 5-Aza-2′-deoxycytidine (5-aza-dC) dissolved in DMSO was obtained from Sigma-Aldrich, USA.