Absence of Rab11a increases the production of defective particles.

Rab11a KO cells show increased production of defective virions. (A-B) Control A549 and Rab11a KO cells were infected with H1N1(PR8) or H3N2 (Pan99) for 48 hours, and supernatants were tittered by plaque assay and subjected to viral RNA extraction. Viral RNAs were reverse transcribed and individual segment copy numbers were quantified by digital droplet PCR. The ratios of viral RNA copy number to PFU are shown for all eight segments of H1N1 and H3N2 viruses. (C) Viral titers at different stages of virion purification. Control A549 and Rab11a KO cells were infected with WT PR8 at MOI = 0.01 and at 72hpi, supernatants were collected and clarified of debris by low speed centrifugation. Subsequently, supernatants were filtered through a 0.45uM filter and pelleted on a 25% sucrose cushion. (D) Comparison of vRNA copy numbers in virions isolated from control A549 and Rab11a KO cells. * p-value < 0.05; ** p-value < 0.01; *** p-value < 0.001; N.S., non-significant. Data shown is a representative of two independent experiments performed with three biological replicates.

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