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. 2023 Feb 14;14:820. doi: 10.1038/s41467-023-36445-9

Fig. 9. APOBEC3B counteracts RNAse L from limiting G3BP1-RNA condensate.

Fig. 9

a U2OS-A3B-flag cells ±DOX were transfected with 2–5 A (20 µM) 8 h after cells were infected with SeV (MOI = 1). At 24hpi, G3BP1, poly(A), and A3B localization were monitored by sequential IF-FISH. Scale bar: 5 μm. b Quantification of poly(A) intensity (arbitrary units) in G3BP1 foci from the experiment described in (a). Red lines indicate the mean (Number of G3BP1 foci, n = 200). P-values were calculated with a two-tailed Welch t-test. c Localization of G3BP1, poly(A), A3B were monitored by sequential IF-FISH after poly(I:C) transfection (200 ng/mL, 16 h) in U2OS-A3B-flag and U2OS-A3B-flag-RNase L KO cells ±DOX. Scale bar: 5 μm. d The surface area of G3BP1 foci was quantified following poly(I:C) (200 ng/mL, 16 h) transfection of the indicated cell lines. Red lines indicate the mean (Number of G3BP1 foci, n = 200). P-values were calculated with a two-tailed Welch t-test. e U2OS cells were knocked down with indicated siRNAs followed by transfection with poly(I:C) (200 ng/mL, 4 h). Poly(A) and G3BP1 localization were monitored by IF-FISH. Scale bar: 5 μm. Quantification of percentage of cells with G3BP1-associated poly(A) in U2OS knocked down for A3B (Number of biological replicates, n = 3) (f) or PABPC1 (Number of biological replicates, n = 4) (g) for 40 h followed by poly(I:C) (200 ng/mL, 4 h) transfection. Mean values ± SD. P-values were calculated with a two-tailed Welch t-test. h Increase of A3B levels promotes G3BP1-RNA condensates in the presence of active RNase L. Source data are provided as a Source Data file.