Ly294002

HL60 and NB4 cells were obtained from the Cell Resource Centre (Xiangya Medical College, Central South University, Hunan, China). The cell lines were maintained in RPMI-1640 medium (Corning Inc., Corning, NY, USA) containing 10% foetal bovine serum (Corning Inc.) and 1% antibiotic solution of penicillin/streptomycin (Sigma, MO, USA) in a 37 °C incubator with a humidified atmosphere of 5% CO₂ (Supplementary Fig. S2). To overexpress GLI1, AML cells were infected with lentivirus containing the GLI1 open reading frame (MOI: 50–100). After 72 h of infection, HL60 and NB4 lines were allowed to recover for 24 h with fresh media and were referred to as HL60/GLI1 and NB4/GLI1 cells (Supplementary Fig. S3). To evaluate the response to drug treatment, 1 × 10⁶ HL60/GLI1 and NB4/GLI1 cells were seeded in 6-well culture plates and incubated with either 20 μM GANT61 (Adooq Bioscience, A13252) or 20 μM LY294002 (Adooq Bioscience, A10547) for 72 h prior to subsequent measurements.

Z-VAD-FMK, chloroquine (CQ), Rapamycin (Rapa), NU7441, Cyt387, A674563, KU60019, LY294002, PD0332991, AT7519, MK2206, CUDC907, LY2109761, GANT61, BIX 02189, Spautin-1, QNZ, LY317615, PD169316, GSK2606414, LYK974, TCK ERK 11e, SC79, MC1568, H89, ICG001, Perifosine, AEE788, ABT263, GDC-0941, FH535, PD0325025, NU7026, STF-083010, AEBSF HCl were purchased from Adooq. N-Acetyl-l-cysteine ethyl ester, DMSO, PEG300, and Tween-80 were purchased from MCE.

MK-2206 (A10003), RAD001 (A10374), CAL-101 (A10172), LY294002 (A10547), and z-VAD-FMK (A12373) were purchased from AdooQ Biosciences. BAY 80-6946 (MedChemExpress, HY-15,346), BYL719 (Cayman Chemical, 16986), NVP-BGT226 (Cayman Chemical, 22142), Staurosporine (Cayman Chemical, 81590), and GSK2126458 (Cayman Chemical, 17377) were purchased from vendors.