Fig. 1.

Activities of TBC1D20 and RN-Tre are important for production of infectious HSV-1. (A) COS7 cells were cotransfected with a plasmid expressing the essential virus protein pUL36 and plasmids expressing TBC domain proteins as described in Materials and Methods. The positive control was transfected with the UL36 plasmid and pcDNA3, while the negative control was transfected with only pcDNA3. At 24 h posttransfection cells were infected with HSV-1ΔUL36 at 5 PFU/cell, progeny virus was collected 16 hpi, and infectious virus levels were determined by titration on HS30 cells. Titers were normalized to the positive control, and bars represent the means of at least three independent experiments. Error bars are standard errors of the means. The solid line represents the median HSV-1 titer across all samples, and dashed lines represent ±1 standard deviation from the median. (B) Rab GAPs of interest (gray bars) were expressed alongside their inactive controls (white bars) and infected as described above. Titers were normalized to the positive control, and bars represent the means of at least two independent experiments. Error bars are standard errors of the means. The dashed line represents a 5-fold reduction in titer compared to the positive control.