1. Fig. 2 BX795 is nontoxic to HCE cells at therapeutic concentration.
BX795 is nontoxic to HCE cells at therapeutic concentration. (A) Apoptosis analysis [Annexin V–FITC (fluorescein isothiocyanate) versus PI] by flow cytometry. HCE cells were incubated with mock, BX795 (10 μM), or TFT (50 μM) for 24 hours and assayed for apoptosis using the Dead Cell Apoptosis Kit. The numbers shown in each quadrant indicate the percentage of cells. The numbers in bold in the top-right and bottom-right quadrants indicate the percentage of early and late apoptotic cells, respectively. Representative plots from three replicates are shown. (B) Cell viability of infected and treated HCE cells assessed by MTT assay. The cells were infected with MOI 0.1 HSV-1(KOS) and, at 2 hpi, treated with mock or BX795 (10 μM). At 24 hpi, viability was assessed by MTT assay. The data (n = 9) are represented as percentage of nontreated, noninfected cells. Significance between the mock-treated and BX795-treated cells was determined by one-way ANOVA with Tukey’s multiple comparisons test. (C and D) Human MAPK phosphorylation array (C) and a heat map (D) depicting the intensity of the spots on the array from HCE cell lysates. The cells were infected with MOI 5 HSV-1(KOS), and at 2 hpi, fresh culture medium containing mock or BX795 (10 μM) treatments was added. At 4 hpi, the cells were collected and the lysates were assayed using the Human MAPK Phosphorylation Kit. Colored boxes indicate the spots on the array, which significantly differ between the mock-treated and BX795-treated cells. Duplicate spots for each sample are shown on the array. (E) Quantification of the kinases shown in (C) relative to the mock-treated cells. Colored bars indicate the kinases that are significantly different. Representative array from two replicates is shown. One-way ANOVA with Dunnett’s multiple comparisons test was performed to determine the significance.
Dinesh Jaishankar et al., Sci Transl Med 2018;10:eaan5861
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