CSAR1 is required for full efficiency of lytic cycle. (A) Western blot stained with anti-FLAG (recognizing ERK7^AID-3xFLAG) and anti-tubulin of lysates from ERK7^AID and Δcsar1 parasites grown in ±IAA. (B–F) Quantification of (B) plaque number, (C) relative plaque size, (D) invasion rate, (E) ionophore-induced egress, and (F) replication rates of ERK7^AID or Δcsar1 parasites grown in ±IAA binned as indicated. B and C also show relative fitness of the wild-type 3xHA-tagged and RING-domain mutant CSAR1 (C1791A/H1792A/H1796A; CSAR1^MUT) parasites. Significance on B–E calculated by 1-way ANOVA followed by Tukey’s multiple comparison test (*** is P < 0.001; n.s., not significant). Data distribution was assumed to be normal, but this was not formally tested. (G) The unbinned cumulative frequency of the data from F. Invasion, egress, and plaque assays are n = 3 biological replicates conducted with n = 3 technical replicates. Replication quantified from three biological replicates of n ≥ 100 vacuoles. Error bars in F are SD, all others are 95% confidence-interval of mean. Replication of Δcsar1 compared to parental ERK7^AID is P < 0.001 as calculated by Kolmogorov-Smirnov test. Source data are available for this figure: SourceData F3.