Posttranscriptional fine-tuning of Ccl3 during mCMV infection. (A and B) Representative contour plots from CCL3-EASER spleen (A) and lung (B) NK cells from mice infected with mCMV and analyzed at the indicated time point and presented in Fig. 4. The red shaded boxes indicate tdTomato-positive cells. (C and D) Representative XY plots of Venus and tdTomato channel intensity from spleen (C) and lung (D) NK cells gated on tdTomato-positive cells (as shown in A and B, respectively). The dotted lines represent the simple linear regression of the correlation between Venus and tdTomato channel intensity. Linear regression is color-coded according to the time point after infection: gray = 0 h (naive), blue = 4 h, green = 8 h, and red = 16 h. (E–G) Representative XY plots of Venus and tdTomato channel intensity from liver (E), spleen (F), and lung (G) monocytes gated on tdTomato-positive cells from mice treated as described in Fig. 4. Linear regression is color-coded as described in C and D. (H) Comparison of simple linear regression from lung monocytes. Color-coded as described in C and D. Black dotted line indicates tdT50: point of half tdTomato channel intensity obtained from the naive group. (I) Frequency and MFI of tdTomato (top panels) and Venus (bottom panels) expression in monocytes during mCMV infection. (J) Viral transcripts determined from liver, spleen, and lungs from the same mice analyzed in E–I. Viral E1 transcripts were quantified by RT-qPCR. The absolute values were normalized to 107 β-actin transcripts. (K) Viral genomes from liver samples in J. Absolute quantification assessed via normalization with 1 × 106 cellular genomes (gB/M55 per 1 × 106PTHrP). Data is presented as mean ± SD; n = 3 mice per group. Statistical analysis was assessed by ordinary one-way ANOVA with Tukey’s post-test to naive group was used: *P < 0.05. In C–G, correlation test was used, two-tailed P value, Pearson r with 95% confidence interval.
