Figure 6. CLAMP MediatesT. gondiiInvasion and Is Essential for theP. falciparumAsexual Cycle
(A) Neighbor-joining tree showing the phylogenetic relationships ofCLAMPhomologs in diverse apicomplexans. Bootstrap values for 10,000 trials are displayed.
(B) Inferred topology of CLAMP highlighting transmembrane domains (orange) and the proline-rich domain (green). See alsoFigure S4.
(C–E) CLAMP-mNeonGreen localization during egress and invasion. Intracellular parasites expressing CLAMP-mNeonGreen were stimulated to egress
with A23187 (C). Relative fluorescence across the length of each parasite (dotted line) is plotted for the four time-points shown. Lines are polynomial regressions±
95% CI (D). The localization was also monitored during invasion (E). The position of the moving junction is indicated with paired open arrows. Solid arrowhead
indicates a punctum of mNeonGreen at the posterior of the parasite appearing immediately after invasion. Time is expressed in minutes:seconds following
addition of the compound (C and D) or initiation of invasion (E). Scale bar, 10mm.
(F) Diagram of the DiCre/CLAMP strain showing how after rapamycin (rapa) treatment the reporter locus switches from expressing KillerRed to expressing YFP,
andCLAMPmRNA degradation is induced.
(G and H) A 2-hr treatment with rapa is sufficient to induceCLAMPdegradation as demonstrated by immunofluorescence microscopy 24 hr later (G) or
immunoblotting 2 days later (H). The parental strain (DiCre) is included as a control.
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