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. 2021 Aug 13;12:4917. doi: 10.1038/s41467-021-25203-4

Fig. 1. Cytoplasmic double-stranded RNAs induce A3A expression by activating RIG-I and MAVS.

Fig. 1

a Indicated DNA (200 ng/mL) or RNA (100 ng/mL) oligonucleotides were transfected in MCF10A cells. A3A mRNA level was monitored by RT-qPCR 16 h after transfection. Mean values ± SD (n = 3). *P < 0.05, **P < 0.01 (two-tailed Welch t-test). Known RIG-I agonist oligonucleotides are labeled in blue. b BICR6 cells were transfected with 3p-hpRNA (40 ng/mL) and A3A mRNA level was monitored at the indicated time. Mean values ± SD (n = 3). c The A3A mRNA levels were monitored in BICR6 cells or indicated knockout cell lines 16 h after 3p-hpRNA transfection. Mean values ± SD (n = 3). **P < 0.01 (two-tailed Welch t-test). d Quantification of DDOST558C>U level in BICR6 WT or RIG-I KO by RNA mutation-based ddPCR assay at the indicated time following 3p-hpRNA transfection (400 ng/mL). Mean values ± SD (n = 4). ****P < 0.0001 (two-tailed Welch t-test). e A3A-deamination activity from indicated cell extracts on a DNA stem-loop substrate containing an ApC motif. f Indicated cell lines were transfected with 3p-hpRNA (100 ng/mL) and Chk1-pS317, A3A, and RIG-I levels were monitored by western blot 24 h after 3p-hpRNA transfection. g BICR6 cells were transfected with A3A siRNA for 48 h followed by 3p-hpRNA transfection (100 ng/mL). A3A and Chk1-pS317 levels were monitored in BICR6 cells 24 h after 3p-hpRNA transfection. h BICR6 WT and RIG-I KO cells were infected with SeV (1 MOI) for 24 h and the A3A mRNA level was quantified by RT-qPCR. Mean values ± SD (n = 3). **P < 0.01 (two-tailed Welch t-test). Source data are provided as a Source Data file.