Extended Data Fig. 7. PP-1-UL32 binding promotes viral growth and the onset of viral gene expression.
(a) Growth curve of mutant and wildtype viruses on HELFs (MOI=5). Means (centre of the error bars) and standard deviations of the mean of n = 3 biological replicates are depicted. Unpaired two-sided t-tests without multiple hypothesis correction were performed comparing the indicated viruses and time points. (b) Kinetics of viral protein levels of wildtype and mutant viruses across the replication cycle (MOI = 5). The experiment was replicated at least three times with similar results. (c) Flow cytometry analysis of Immediate Early (IE) protein levels as a function of the cell cycle stage. HELFs were infected with the indicated recombinant viruses (MOI = 5, 6 h post infection). The percentage of IE1/2-positive cells in G1 or S/G2 compartments is given. (d, e) Western blot analysis of abundance levels of selected proteins of purified virions for RxLmut and RxL/SILK/RVxFmut viruses. Exemplary blots in (d) and quantification based on n=4 (14-3-3) or n=3 (PP-1, UL32) technical replicates in (e). The height of the bars corresponds to the mean and the error bars to the standard deviation. P-values based on two-sided t-tests without multiple hypothesis correction. Control experiments to Fig. 5b.