Fig. 2: Cell cycle or size cannot explain variation in HSV-1 transcription or translation.
From: Cellular state landscape and herpes simplex virus type 1 infection progression are connected
a PCNA, ICP27, and ICP8 staining of HSV-1-infected HeLa cells at 12 hpi. Arrowheads, ICP27 expressing cells. Arrow, an infected cell containing viral RCs with co-localized PCNA. Nucleus and cell outlines as in Fig. 1a. Scale bar, 25 μm. b Mean nuclear intensity of PCNA in uninfected or infected HeLa cells from the smFISH + 4i experiment. Data are from four individual replicate wells per time point (1.5 hpi, nuninf. = 18,442 and ninf = 8665; 3 hpi, nuninf. = 26,732 and ninf = 2744; 6 hpi, nuninf. = 22,558 and ninf = 7537; 9 hpi, nuninf. = 28,858 and ninf = 9382; 12 hpi, nuninf. = 25,851 and ninf = 9914 cells). PCNA distributions were compared using pairwise two-sided Kolmogorov–Smirnov (KS) test: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns not significant. Boxplots definitions as in Fig. 1c. Points, outliers. c Cell-cycle phase of uninfected or infected HeLa cells from the smFISH + 4i experiment. Data are from three or four individual replicate wells per time point (1.5 hpi, nuninf. = 18,442 and ninf = 8665; 3 hpi, nuninf. = 26,732 and ninf = 2744; 6 hpi, nuninf. = 22,558 and ninf = 7537; 9 hpi, nuninf. = 21,920 and ninf = 7155; 12 hpi, nuninf. = 25,851 and ninf = 9914 cells). Statistical comparison is shown in Supplementary Fig. 4b. d Single-cell UL29 and UL19 transcript counts and ICP27 and ICP4 mean intensities in infected G1, S, and G2-phase HeLa cells. UL29 and UL19 transcript counts per cell were normalized by dividing the counts by the cell size and multiplying by 1000. 0.1–99.9th percentiles of marker intensities are shown in the density plots. Data are from three or four individual replicate wells per time point from the smFISH + 4i experiment. Inset: cell counts per time point. e Single-cell UL29 and UL19 transcript counts and ICP27 and ICP4 sum intensities versus cell size (UL29, n = 20,225; UL19, n = 9045; ICP27 and ICP4, n = 29,577 cells). Data are from four individual replicate wells per time point from the smFISH + 4i experiment. Inset: R2 of robust linear regression models fit to the single-cell data per time point. See also Supplementary Figs. 4 and 5.
