Fig. 6: HSV-1 activates Akt to suppress apoptosis and to enhance translation.
From: Cellular state landscape and herpes simplex virus type 1 infection progression are connected
a Cleaved caspase 3 as a function of p-Akt (Ser473) in the infected HeLa cells at 12 hpi from the smFISH + 4i experiment (n = 4 wells). b HeLa cells were infected with HSV-1 (MOI 1) and p-Akt and ICPs were detected in untreated and DMSO- or LY294002-treated cells at 12 hpi using Western blotting. Treated samples were compared to the untreated samples to give a fold change. Data represent mean ± standard deviation from three independent experiments. Treatments were compared by one-way ANOVA with Tukey’s multiple comparison post-test relative to untreated samples. c Single-cell intensity of 4E-BP in mock or HSV-1-infected in HeLa cells after DMSO or LY294002 treatment, from three replicate wells (mock nDMSO = 34,389, nLY294002 = 23,533; HSV-1 MOI 0.3, nDMSO = 28,067, nLY294002 = 19,455; HSV-1 MOI 1 nDMSO = 25,360, nLY294002 = 20,700 cells). 4E-BP and ICP27 were detected by immunofluorescence imaging at 12 hpi. Right: mock cells (0) and ICP27-expressing cells from HSV-1 infection using MOI 0.3 (3). The overlapped estimated area of two distributions is indicated as a mean percentage from two biologically independent experiments (n = 3 wells per experiment). Data from a biological replicate are shown in Supplementary Fig. 13b. d Apoptotic cells in mock or HSV-1-infected HeLa cells (MOI 0.3) treated with DMSO or LY294002. Cleaved caspase 3 and ICP27 were detected by immunofluorescence imaging at 12 hpi. Mock cells (nDMSO = 32,328 and nLY294002 = 24,073 cells) and ICP27-expressing cells from HSV-1-infection (nDMSO = 13,376 and nLY294002 = 10,513 cells). Data are from one experiment and represent mean ± standard deviation among three replicate wells (Supplementary Fig. 13c shows data from a biologically independent experiment). Percentage of apoptotic cells was compared between the treatments by one-sided unpaired two-sample t-test. e Mean intensity of cleaved caspase 3 as a function of ICP27 in the LY294002-treated, ICP27-expressing HeLa cells (nnon-apoptotic = 8713 and napoptotic = 1800 cells). Data are from the same experiment as in d. In b and d: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. 0.1–99.9th percentiles of marker intensities are shown in the density plots. See also Supplementary Fig. 13.
