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Figure 2 | BMC Biotechnology

Figure 2

From: High level expression of soluble glycoproteins in the allantoic fluid of embryonated chicken eggs using a Sendai virus minigenome system

Figure 2

Analysis of soluble proteins in allantoic fluids. (A) Allantoic fluids (4.5 ml) from eggs inoculated with mixed stocks of SeV and SeV minigenomes expressing HA-M, HRSV/FTM- or HMPV/FTM- were centrifuged through a 25% glycerol cushion. Pellets were resuspended in 100 μl of PBS. All fractions were analyzed by Western blot. Five microliters of allantoic fluid from uninfected eggs (0) were compared with 5 μl of allantoic fluid from eggs inoculated with virus stocks encoding the indicated proteins (1), 5 μl of resuspended pellet (2), and 15 μl of the ultracentrifugation supernatant (3). (B) Comparison by Western blot of the expression levels of HRSV/FTM-and HMPV/FTM- from allantoic fluid or the supernatant of rVV-infected cells. Twenty microliters of culture supernatant from mock-infected HEp-2 cells (1), and 5, 10, or 20 μl of supernatant from rVV-infected cells (2, 3 and 4, respectively) were compared with 20 μl of uninfected allantoic fluid (5) and 5, 10, or 20 μl of allantoic fluid harvested from passage 4 (see Figure 1C) of defective SeV virus-inoculated embryonated eggs (6, 7 and 8, respectively). Asterisk (*) denotes a rVV band that cross-reacts with anti-F specific reagents.

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