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. 2017 Mar 15;2(2):e00054-17. doi: 10.1128/mSphere.00054-17

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Copyright © 2017 Huang et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 3 — Complementation of the AP2IX-4 knockout. (A) Schematic illustrating the generation of the complemented knockout (Δap2IX-4::AP2IX-4^HA). A cDNA-derived version of AP2IX-4 tagged at the C terminus with 3xHA (complementation construct) was targeted to the ablated genomic locus in Δap2IX-4 parasites. (B) Genomic PCRs were performed with the indicated primer pairs to confirm proper integration of the complementation construct into Δap2IX-4 parasites. Primers corresponding to GAPDH were used as a positive control of the DNA preparation. (C) Western blot of Δap2IX-4 and Δap2IX-4::AP2IX-4^HA parasites with anti-HA, using anti-β-tubulin as a control to verify the presence of parasite protein. (D) IFAs were performed on Δap2IX-4::AP2IX-4^HA tachyzoites as described for Fig. 1C. (E) Quantification of the proportion of parasite vacuoles (of 100) in the asynchronous population of AP2IX-4^HA parasites performed as described for Fig. 1D.