Extended Data Fig. 2: Characterization of mScarlet reporter YFVs and fluorescence activated cell sorting (FACS) results. | Nature Microbiology

Extended Data Fig. 2: Characterization of mScarlet reporter YFVs and fluorescence activated cell sorting (FACS) results.

From: Amino acid changes in two viral proteins drive attenuation of the yellow fever 17D vaccine

Extended Data Fig. 2

a, Genome organization of full-length YFV (upper) and the mScarlet reporter YFV (bottom). A gene cassette containing 11 amino acids of the NS1 N-terminus for protein processing, an mScarlet reporter gene, and a gene encoding for the transmembrane domain of the dengue virus envelope protein (DENV-linker) were inserted in-frame between YFV E and NS1. b, mScarlet-positive cells were measured at 48 hpi. by flow cytometry at indicated MOIs. Data are mean ± s.d. from 1 experiment with 3 biological samples (n = 3). c, Gating strategy of mScarlet-positive cells for collection by FACS and subsequent RNA-Seq. mScarlet-negative cells from mock infected Huh7 cells and high mScarlet-expressing cells from infected Huh7 cells were sorted for RNA-Seq. The individual sorting results of each sample are shown. d, YFV genome copy numbers were measured by RT-qPCR from total RNA of sorted cells. LOD, limit of detection. Data are mean ± s.d. from 6 experiments with 1 biological sample (n = 6). P values were calculated by two-way (b) or one-way (d) ANOVA with Tukey ad hoc test for multiple comparison.

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