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Figure 1. RCN1 knockdown is associated with cGAS-STING signaling in AML cells. (A) Pyroptotic cell death was assessed in NB4 cells transfected with shNC (n = 3) or shRCN1 lentiviruses (n = 3), with or without STING-I treatment. Measurements were performed on day 5 via the detection of active caspase-1 using a FLICA® 660 assay; (B) Cellularity of NB4 cells transfected with shNC (n = 3) or shRCN1 (n = 3) lentivirus and treated with or without STING-I on day 5; (C and D) Cellularity of THP-1 (C) and OCI-AML3 (D) cells following RCN1 deficiency; (E-G) Western blot detection of key proteins in the cGAS-STING-type I interferon pathway (including IFN-β and OAS3) in NB4 (E), THP-1 (F), and OCI-AML3 (G) cells treated with shNC or shRCN1 lentiviruses; (H-J) The mRNA expression levels of ISGs including IFIT1, IFI27, RSAD2, OAS1, and ISG15 in RCN1-deficient NB4 (H), THP-1 (I), and OCI-AML3 (J) cells, as determined by qPCR. Results are shown as mean ± standard deviation. Statistical analyses were performed using one-way ANOVA followed by Tukey’s multiple comparisons test for panels involving more than two groups (A and B) and a two-tailed unpaired t-test for panels involving two-group comparisons (C and D, H and J). ^*P < 0.05; ^**P < 0.01; ^***P < 0.001; ^****P < 0.0001. AML: Acute myeloid leukemia; ANOVA: analysis of variance; cGAS: cyclic GMP-AMP synthase; FLICA: fluorescent-labeled inhibitor of caspases; IFI27: interferon alpha-inducible protein 27; IFIT1: interferon-induced protein with tetratricopeptide repeats 1; IFN-β: interferon beta; ISG15: interferon-stimulated gene 15; ISGs: interferon-stimulated genes; mRNA: messenger RNA; OAS1: 2’-5’-oligoadenylate synthetase 1; OAS3: 2’-5’-oligoadenylate synthetase 3; qPCR: quantitative polymerase chain reaction; RCN1: reticulocalbin 1; RSAD2: radical S-adenosyl methionine domain containing 2; shNC: short hairpin negative control; shRCN1: short hairpin RNA targeting RCN1; STING: stimulator of interferon genes; STING-I: STING inhibitor.