PfHO localization and processing. A) Widefield fluorescence microscopy of Dd2 parasites episomally expressing PfHO-GFP. For live imaging, parasites were stained with 25 nM Mitotracker Red and 10 nM Hoechst. For IFA, parasites were fixed and stained with anti-GFP and anti-apicoplast ACP antibodies, as well as DAPI. For all images, the white scale bars indicate 1 μm. The average Pearson correlation coefficient (rp) of red and green channels based on all images is given. Pixel intensity plots as a function of distance along the white line in merged images are displayed graphically beside each parasite. B) Western blot of untreated or Dox/IPP-treated parasites episomally expressing PfHO-GFP and stained with anti-GFP antibody. C) Live microscopy of PfHO-GFP parasites cultured 7 days in 1 μM doxycycline (Dox) and 200 μM IPP and stained with 25 nM Mitotracker Red and 10 nM Hoechst. D) Widefield fluorescence microscopy of Dd2 parasites episomally expressing PfHO N-term-GFP and stained as in panel A. E) Western blot of parasites episomally expressing PfHO N-term-GFP and stained with anti-GFP antibody. F) Live microscopy of PfHO N-term-GFP parasites cultured 7 days in 1 μM Dox and 200 μM IPP, and stained as in panel C. For each parasite line and condition, ≥20 parasites were analyzed by live imaging and ≥10 parasites were analyzed by IFA. Figure supplement 1. Additional widefield fluorescence microscopy of live Dd2 parasites episomally expressing PfHO-GFP and PfHO N-Term-GFP. Figure supplement 2. Additional widefield IFA microscopy of fixed Dd2 parasites episomally expressing PfHO-GFP and PfHO N-Term-GFP. Source data 1. Uncropped western blots of untreated or Dox/IPP-treated parasites episomally expressing PfHO-GFP in figure 2B. Source data 2. Uncropped western blots of untreated parasites episomally expressing PfHO N-term-GFP in figure 2E.Blackwell AM, Jami-Alahmadi Y, Nasamu AS, Kudo S, Senoo A, Slam C, Tsumoto K, Wohlschlegel JA, Caaveiro JMM, Goldberg DE, Sigala PA. Malaria parasites require a divergent heme oxygenase for apicoplast gene expression and biogenesis. bioRxiv [Preprint]. 2024. PMID: 38853871

È