(A) Size distribution and percentage of AGO-bound reads mapped to influenza following infection of BMDMs with influenza or influenza del NS1 (MOI 1, 24h.p.i).
(B) Immunoblot of pan-AGO or AGO2 of cell lysates or (C) pan-AGO of immunoprecipitates.
(D) Amount of AGO-loaded influenza RNA in influenza-infected BMDMs, relative to WT controls.
(E) Quantity of AGO-loaded influenza-derived RNA at indicated nucleotide lengths.
(F) Origin of AGO-bound RNA that mapped to the influenza A genome. Genomic segments presented from the 3′ end (left) to the 5′ end (right), and negative (−) or positive (+) strand and scale are indicated. Boxes highlight bona fide vsiRNAs (Li et al., 2016; Qiu et al., 2017).
(G) Percentage and size distribution of AGO co-immunoprecipitated influenza RNA in BMDMs infected with influenza delNS1 (MOI 1, 24 h p.i.).
(H) Relative influenza HA RNA as quantified by qPCR in MEFs infected with influenza or influenza delNS1 in the presence or absence of 10 μg/mL anti-IFNAR.
Data are the mean of two or three biological replicates. Error bars represent SEM. *p < 0.05, **p < 0.01, and ***p < 0.001, as determined by an unpaired t test or one-way ANOVA with a Bonferroni multiple comparisons test.