1. Fig. 1 BX795 suppresses HSV-1 infection.
BX795 suppresses HSV-1 infection. (A and B) HCE cells were infected with multiplicity of infection (MOI) 1 HSV-1(K26RFP) with BX795 at the indicated concentrations for 2 hours, and then fresh culture medium containing BX795 was added. Mock treatment was used as a control. Virus yields were determined at 24 hours post-infection (hpi) using flow cytometry (A) [represented as percentage of mock-treated mean fluorescence intensity (MFI)] and qPCR (B) (represented as percentage of mock-infected cells) (n = 3 replicates). (C) Representative micrographs (left panel) of HCE cells depicting the presence of HSV-1 (green). HCE cells were infected with HSV-1(K26GFP) at the indicated MOIs, and at 2 hpi, fresh culture medium containing mock, TFT (50 μM), or BX795 (10 μM) was added to the cells. Images were captured at 24 hpi using AxioVision 100 under 10× objectives. Scale bar, 100 μm. The number of GFP-infected cells per field was calculated using the MetaMorph imaging software (right panels, n = 3 fields). Data are represented as percentage of mock-infected cells. Significance to mock-treated cells was determined by one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons test. (D) HSV-1 VP16 and gD transcripts determined by qRT-PCR. Infection was performed as mentioned in (C). Cells were collected at 24 hpi to extract RNA, and viral transcripts were quantified by qRT-PCR and represented as fold change over the mock-treated cells. Significance to mock-treated cells was determined by one-way ANOVA followed by Dunnett’s multiple comparisons test (n = 4 replicates). (E) Immunoblots of HSV-1 gB protein and human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from infected HCE cells at 24 hpi. Infection was performed as mentioned in (C). Representative blots from three replicates are shown.
Dinesh Jaishankar et al., Sci Transl Med 2018;10:eaan5861
Published by AAAS