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

Fig. 2. PABPC1 recruits APOBEC3B to stress granules.

Fig. 2

a A3B pulldown strategy. b Indicated pulldown samples from HEK-293T cells were analyzed by SDS-PAGE and silver staining. c DOX-treated U2OS-A3B-flag cells were infected with SeV (MOI = 1) and the cellular localization of the indicated proteins was monitored by immunofluorescence at 24hpi. Scale bar: 5 μm. d GFP pulldown of A3B-GFP/Flag or A3B-ΔNTD-GFP/Flag expressed in HEK-293T cells were analyzed by western blot with the indicated antibodies. e Immunoprecipitation of PABPC1 from U2OS-A3B-flag cells ± DOX and analyzed by western blot with the indicated antibodies. f Quantification of A3B intensity (arbitrary units) in G3BP1 foci in U2OS-A3B-flag cells treated with DOX and transfected with a siRNA control (CTL) or against PABPC1 for 40 h and then treated with NaAsO2 (500 µM, 1 h). Red lines indicate the mean (Number of G3BP1 foci, n = 150). P-values were calculated with a two-tailed Welch t-test. g Immunofluorescence for A3B (Flag) and G3BP1 in U2OS-A3B-flag cells + DOX transfected with siCTL or siPABPC1 for 40 h and treated with NaAsO2 (500 µM, 1 h). Scale bar: 5 μm. h Quantification of A3B intensity (arbitrary units) in G3BP1 foci in U2OS-A3B-flag cells treated with DOX, transfected with a siRNA control (CTL) or against PABPC1 for 40 h, and then transfected with poly(I:C) (200 ng/mL, 16 h). Red lines indicate the mean (Number of G3BP1 foci, n = 150). P-values were calculated with a two-tailed Welch t-test. i Quantification of the number of cytoplasmic G3BP1 foci by cell in individual U2OS cells knocked down for PABPC1 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. Source data are provided as a Source Data file.