Figure 3.

ZAPS and ZAPL binding to alphavirus genomic RNA do not correlate with alphavirus replication sensitivity to ZAP. (A,C) Lysates of ZAP KO 293T cells with induced expression of RFP-ZAPS (A) or RFP-ZAPL (C) were incubated with biotinylated Fluc RNA or gRNA of RRV, SINV, ONNV, or CHIKV. Following streptavidin-mediated pull-down of biotinylated RNA, ZAP associated with each RNA and in whole cell lysate (input) was assayed by immunoblot (IB). Expected sizes: RFP-ZAPS = 106 kDa, RFP-ZAPL = 130 kDa. Data are representative of results from three independent experiments. (B,D) ImageJ quantifications of panel A (B) or C (D). Data are combined from three independent experiments. Error bars represent the SD. Asterisks indicate statistically significant differences (one-way ANOVA and Tukey’s multiple comparisons test: *, p < 0.05; **, p < 0.01; ****, p < 0.0001).