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. 2019 Dec 4;10:5518. doi: 10.1038/s41467-019-13520-8

Fig. 6. The 3′ splice site is a host range determinant.

Fig. 6

a (Top) Eight nucleotide polymorphisms of Mal virus at the M segment 3′ splice site were integrated into wild-type Pan virus generating “Pan-Av” mutant. a (Bottom) Design of pulsed SILAC experiment. A549 cells were infected with wild-type or mutant virus at an MOI of 4 (FFU/cell) and viral gene expression was assessed by pulse labeling during two time intervals. b pSILAC-based quantification of viral protein production comparatively for Pan and Pan-Av virus. One replicate (n = 1) is depicted. c Kinetics of M1/M2 protein levels as detected by immunoblotting. d RNA-seq-based quantification of viral mRNA levels at 8 hpi (MOI = 1 FFU/cell) comparing Pan and Pan-AV infection. The average of n = 2 biological replicates is depicted. e Quantification of M segment-derived RNAs based on splice junction reads. The area of the pies scales with the total number of splice junction reads. The average of n = 2 biological replicates is depicted. f Multicycle replication curve of the indicated viruses at an MOI of 0.05 FFU/cell on A549 cells. Means and standard deviations of biological triplicates (n = 3) are shown along with significance estimates based on two-sided paired t tests for the 16–48 h time points (n.s.: non-significant, **p < 0.01, ***p < 0.001).