Aim2 1

HL-60 and THP-1 cells were infected with MISSION lentivirus carrying pLKO.1 puro with shRNA constructs (STING-1: TRCN0000164628; STING2: TRCN0000135555; AIM2-1: TRCN0000107501; AIM2-2: TRCN0000107502; and cGAS: TRCN0000149984), which were purchased from Sigma-Aldrich. Each cell line infected with MISSION lentivirus carrying pLKO.1 puro and with a non-mammalian shRNA control construct was used as a shControl. Stably transfected cells were selected in culture medium supplemented with 1 μg/ml puromycin for more than 2 weeks. For the transient KD of ATG5 gene expression, HL-60 cells were transfected with ATG5 siRNA (Santa Cruz Biotechnology, Santa Cruz, CA, USA) using GENOMONE-Si transfection reagent (Ishihara Sangyo, Osaka, Japan). As a siControl, cells were transfected with control siRNA-A (Santa Cruz Biotechnology). The KD efficiency of each gene was confirmed using quantitative real-time PCR (qRT-PCR) and western blotting analyses.

STING and AIM2 knockdown (KD) cells were previously established from HL-60 or THP-1 cell lines, and their KD efficiencies were determined (27 (link)): shSTING-1 HL-60, 83.1%; shSTING-2 HL-60, 83.3%; shAIM2-1 HL-60, 93.1%; shAIM2-2 HL-60, 91.8%; shAIM2-1 THP-1, 73.1%; shAIM2-2 THP-1, 60.1%. AIM2 KD MOLM-14 cell lines were newly established by infection with MISSION lentivirus carrying pLKO.1 puro with short hairpin RNA (shRNA) constructs (AIM2-1: TRCN0000107503, AIM2-2: TRCN0000107504), which were purchased from Sigma-Aldrich. To establish p21 KD cells, HL-60 cells were infected with MISSION lentivirus carrying pLKO.1 puro with shRNA constructs (p21-1: TRCN0000040123, p21-2: TRCN0000287021). As a control, each cell line was infected with MISSION lentivirus carrying pLKO.1 puro with non-mammalian shRNA control constructs. Stably transfected cells were selected in culture medium supplemented with 1 µg/mL puromycin for more than 2 weeks.