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. 2020 Feb 11;15(2):e0228572. doi: 10.1371/journal.pone.0228572

Fig 9. Serial passage of HPIV3-RSV F vectors in Vero cells: Analysis of the stability of RSV F expression.

Fig 9

The P4 stock of F-H3TMCT/N-P, P2 stock of F/preN, F-H3TMCT/preN, and F/N-P, or wt rHPIV3 was passaged serially five times in Vero cells, with four replicates of each. Virus samples from each passage were analyzed by double-staining plaque assay on Vero cells to measure the stability of RSV F expression. A. Scans of the immunostained monolayers made with the Odyssey infrared imaging system (Licor) using 800-nm and 680-nm channels: HPIV3 plaques expressing RSV appear as yellow, while HPIV3 plaques in which expression of RSV F was silenced appear as green. Large plaque variants also are evident in some wells. B. Green plaques indicating loss of RSV F expression, and large plaque variants, were counted and are shown as mean % plaques with loss of RSV F expression and mean % large plaques.