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. 2018 Feb 22;14(2):e1006836. doi: 10.1371/journal.ppat.1006836

Fig 4. TgATrx1 and TgATrx2 are essential and their function requires their CXXC motifs.

Fig 4

A. Scheme of the manipulation performed to each of the TgATrxs loci to replace their promoters with the tetracycline-regulatable promoter. The gene model in the scheme is based on TgATrx1’s gene model, however the promoter replacement occurred via the same strategy for both TgATrxs. Black boxes–exons; asterisk–stop codon. B. Western blot analysis of TgATrx1 expression using anti-TgATrx1 (i), and endogenously HA-tagged TgATrx2 using anti-HA (ii), upon ATc treatment. C. Fluorescent microscopy showing TgATrx1 (anti-TgATrx1, bottom, green) and TgATrx2 (anti-HA, top, green) depletion at 48 hours of ATc treatment (+ATc) compared to non-treated control (-ATc). D. Plaque assays performed with TATiΔKu80PIATrx1 (top) and TATiΔKu80PIATrx2-3HA (bottom) with (+) or without (-) ATc. E. Plaque assays performed with TATiΔKu80PIATrx1 constitutively expressing a copy of TgATrx1CXXC (i) or TgATrx1CXXA (ii) and with TATiΔKu80PIATrx2-3HA constitutively expressing a copy of TgATrx2CXXC (iii) or TgATrx2CXXA (iv). F. Fluorescent microscopy of the localization of TgATrx1CXXC (i); TgATrx1CXXA (ii); TgATrx2CXXC (iii) and TgATrx2CXXA (iv), all in green, co-stained with Streptavidin (SA) which labels the apicoplast acetyl CoA carboxylase (i, ii) or CPN60 (iii, iv) both in red. White broken line shows parasites’ shapes. Scale bar, 1 μm.