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. 2021 Jul 6;17(7):e1009689. doi: 10.1371/journal.ppat.1009689

Fig 2. Growth of VZV containing DHFR degron sequence on specific genes confers TMP-conditional virus growth and formation of plaques.

Fig 2

(A-C) Conditional growth analysis of three VZV after initiating the infection of monolayers at low multiplicity with VZV degron virus or pOka. Cultures were grown in the presence or absence of 100 nM TMP maintained throughout the growth period from 0–4 days post-infection (dpi). Each day, cultures were then trypsinized and the infectious cell titer was determined by growth on new monolayers in conditions in which TMP was provided in the media. Shown are growth analyses for (A) VZV ORF9cDHFR, (B) VZV ORF4nDHFR, and (C) VZV ORF63cDHFR. (D) Plaque size of wild-type VZV (pOka) or VZV derived from the parental BAC (parent) or containing the degron addition to the respective genes compared under permissive (100 nM TMP, blue) or nonpermissive (0 nM TMP, red) conditions at 4-dpi. Images (n = 28–35) were acquired for each virus/condition under identical settings and evaluated as detailed in methods and presented in pixel size estimated from images acquired under 10X (NA.030) air objective. Error bars: SD.