Conditional disruption of the sub2 gene within a single erythrocytic cycle.(B) IFA localising SUB2HA3 in SUB2HA3:loxP schizonts. Parasite nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI, blue). Scale bar, 10 mm.. Immunofluorescence analysis (IFA) of two integrant clones of the modified parasite line (called SUB2HA3:loxP) with anti-HA3 antibodies showed a signal consistent with the previously-determined location of SUB2 in micronemes, confirming correct gene modification. (E) IFA showing loss of SUB2HA3 in cycle 0 SUB2HA3:loxP schizonts following RAP treatment of rings. Expression was detectable in just 2 ± 0.1% of RAP-treated schizonts (the microscopic field was deliberately chosen to include 2 of the rare HA3-positive schizonts). No signal was detectable by IFA in most of the RAP-treated schizonts.

Collins CR, Hackett F, Howell SA, Snijders AP, Russell MR, Collinson LM, Blackman MJ. The malaria parasite sheddase SUB2 governs host red blood cell membrane sealing at invasion. Elife. 2020 9:e61121.. PMID: 33287958