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

Fig. 3. APOBEC3B promotes PKR activity during viral infections.

Fig. 3

a Quantification of the number of G3BP1 foci by cell in U2OS-A3B-flag cells ± DOX and infected with SeV (MOI = 1, 24hpi). Top; percentage of cells with G3BP1 foci. Red lines indicate the mean (Number of cells, n = 500). P-values were calculated with a two-tailed Welch t-test. b, c U2OS or A549 cells were transfected with siRNA against A3B for 40 h and subsequently infected with SeV at MOI = 1. At 24hpi, the levels of G3BP1 foci were quantified by immunofluorescence. Top; percentage of cells with G3BP1 foci. Red lines indicate the mean (Number of cells, n = 500). P-values were calculated with a two-tailed Welch t-test. d Quantification of the number of G3BP1 foci in U2OS KO A3B cells infected with SeV (MOI = 1, 24hpi). Top; percentage of cells with G3BP1 foci. Red lines indicate the mean (Number of cells, n = 500). P-values were calculated with a two-tailed Welch t-test. e U2OS-A3B-flag cells ± DOX were transfected with a siRNA targeting the 3′UTR of endogenous A3B mRNA or a control (siCTL) for 40 h. The levels of G3BP1 foci were quantified by immunofluorescence 24hpi with SeV at MOI = 1. Top; percentage of cells with G3BP1 foci. Red lines indicate the mean (Number of cells, n = 500). P-values were calculated with a two-tailed Welch t-test. f U2OS-A3B-flag cells ± DOX were infected with SeV (MOI = 1, 24hpi) and analyzed by western blot with antibodies against the indicated proteins. U2OS cells knocked down (g) or knocked out (h) for A3B were infected with SeV (MOI = 1, 24hpi) and analyzed by western blotting using the indicated antibodies. i U2OS-A3B-flag cells ± DOX were transfected with an siRNA targeting the 3′UTR of endogenous A3B mRNA or a control (siCTL) for 40 h. The levels of PKR-pT446, A3B, and GAPDH were detected by western blot 24hpi with SeV at MOI = 1. U2OS cells were transfected with siA3B for 40 h and then infected with either Poliovirus (PV) at MOI = 1 for 20 h (j), infected with Sindbis Virus (SINV) at MOI = 1 for 16 h (k), or transfected with 200 ng/mL poly(I:C) for 8 h (l). The levels of PKR-pT446, A3B, and Vinculin were analyzed by western blotting. Source data are provided as a Source Data file.