fig4

Figure 4. TFAM mediates cGAS-STING pathway activation induced by RCN1 deficiency. (A and B) Cellularity of NB4 (A) or OCI-AML3 (B) cells infected with shNC (n = 3) or shTFAM (n = 3) lentiviruses; (C and D) Relative cytosolic mtDNA levels in NB4 (C) or OCI-AML3 (D) cells infected with shNC (n = 3) or shTFAM (n = 3) lentiviruses measured by qPCR; (E and F) Expression of cGAS-STING pathway proteins, IFN-β, and OAS3 in TFAM-knockdown NB4 (E) or OCI-AML3 (F) cells; (G) Changes in cellularity after TFAM overexpression in RCN1-deficient NB4 cells; (H) Cytosolic mtDNA levels in RCN1-deficient NB4 cells following TFAM overexpression were analyzed by qPCR; (I) Expression of cGAS-STING-related proteins in RCN1-deficient NB4 cells following TFAM upregulation; (J) Impact of TFAM overexpression on ISGs (OAS1, IFIT1, IFI27, RSAD2, and ISG15) mRNA levels in RCN1-knockdown NB4 cells measured via qPCR. Data are presented as mean ± standard deviation. Statistical significance was assessed using a two-tailed unpaired t-test. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001; ^****P < 0.0001. cGAS: Cyclic GMP-AMP synthase; 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; mtDNA: mitochondrial DNA; 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; shTFAM: short hairpin RNA targeting TFAM; STING: stimulator of interferon genes; TFAM: mitochondrial transcription factor A.