Fig. 2: TgLIPIN is indispensable for parasite replication and growth within its host.

a Western blot shows TgLIPIN downregulation, 48 h + ATc (0.5 μg/mL), TgLIPIN (anti-HA), and TOM40 (control). b IFA of TgLIPIN-ikD, indicating loss of protein using anti-HA antibody at 48 h +ATc, Scale bar: 2.0 μm. c Replication rate of TgLIPIN parasites grown with (+) or without (−) ATc measured by parasite number per parasitophorous vacuole after 24 h of growth post infection (>100 vacuoles were counted per biological replicate; n = 3, unpaired t test P values where **P = 0.002 −ATc vs +ATc 1 parasite/vacuole, ****P < 0.000001 −ATc vs +ATc 2/4-6/8-10 and abnormal parasite/vacuole). Data are presented as mean values + /− SEM. d TgLIPIN-ikD plaque assays measuring parasite growth over 8–10 days (+/−ATc) in different FBS conditions. e shown as a bar graph (10%, 1% and 0%; n = 4, unpaired t test P values where ****P < 0.000001 −ATc vs +ATc 10/1/0% FBS, **P = 0.026 +ATc 10% FBS vs +ATc 1% FBS, ***p = 0.0001 +ATc 10%FBS vs +ATc 0% FBS, **P = 0.0047 +ATc 1% FBS vs +ATc 0% FBS). Data are presented as mean values + /− SEM. f IFA illustrating early phenotypic effects of TgLIPIN depletion (24 h+ ATc) showing the presence of residual protein (HA-green). Inner membrane complex antibody (IMC- red) clearly shows aberrant IMC membrane biogenesis. g IFA illustrating the phenotypic effects of TgLIPIN depletion after 48 h (+ATc) using anti-IMC antibody in TgLIPIN-ikD. Scale bar: 5.0 μm.