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

Fig 3. Proteins produced by VZV ORF9cDHFR and pOka grown under media containing or without TMP.

Fig 3

(A) Cells were infected with titrated virus infected cell stocks as detailed in methods to allow multi-step virus growth analysis with VZV ORF9cDHFR or pOka. Proteins in infected cultures were harvested at times 24, 48 or 72-hpi in the presence (+) and absence (-) of 100 nM trimethoprim (TMP) and compared to uninfected cell extracts. SDS PAGE separated proteins were immunoblotted and probed with antibodies to ORF9, or proteins from other herpesvirus kinetic classes (IE62, ORF29p), and a cellular control (alpha-tubulin). Signals were determined using a LICOR Odyssey IR imager. ORF9 shows multiple species due to several recognized phosphorylated forms, and the expected size increase (~18 kDa) due to degron motif addition. The sizes of marker proteins (kDa) are indicated to the left of the respective blots. The blots are scanned in linear range and are representative of two identical experiments.