Fig. 2

UBLCP1 is essential for parasite growth in Toxoplasma gondii. A, Schematic illustration showing the replacement of the coding sequence of UBLCP1 using a CRISPR–Cas9 method. B, PCR amplification of UBLCP1 fragment in RHΔku80 and mutant strains. PCR1 indicates the absence of UBLCP1; PCR2 and PCR3 indicate the 5′ and 3′ integration of homologous fragments, respectively. C, Relative mRNA levels of UBLCP1 in the parental RHΔku80 and ΔUBLCP1 strains of T. gondii. Data are presented as the mean ± standard deviation (SD) of three independent repeats. ****P ≤ 0.0001, using a t-test. D, Representative image of plaques observed in HFFs infected with the parental RHΔku80 or ΔUBLCP1 strain 7 or 12 days postinfection. E, The relative number and area of plaques formed by tachyzoites of indicated strains 7 days postinfection. F, The number of attached and invaded parasites of RHΔku80 or ΔUBLCP1 tachyzoites to/into HFFs. G, The invasion efficiency of RHΔku80 and ΔUBLCP1 tachyzoites, presented as invaded parasites/total parasites. H, The intracellular replication rates of RHΔku80 and ΔUBLCP1 are indicated by the number of tachyzoites per parasitophorous vacuole (PV). At least 100 vacuoles are analyzed for each mutant