Plan apochromat 100 1.4 oil objective

All fluorescence images were captured using a Zeiss Axioskop 2plus microscope with a Hamamatsu digital camera (model C4742-95) or a Leica D6B fluorescence microscope equipped with a Leica DFC9000 GT camera and a Leica Plan Apochromat 100×/1.4 oil objective.
Microscopy of live-parasite-infected erythrocytes was performed as previously described (68 (link)). Briefly, parasites were incubated in standard culture medium with 1 μg/ml Hoechst-33342 (Invitrogen) for 15 min at 37°C prior to imaging. Infected erythrocytes (5.4 μl) were added on a glass slide and covered with a cover slip. Nuclei were stained with 1 μg/ml Hoechst-33342 (Invitrogen). Mitochondria were visualized by incubation of parasites with 20 nM MitoTracker Red 665 CMXRos (Invitrogen) for 15 min at 37°C prior to imaging. Contrast and intensities were linear adjusted for clarification and cropped images were assembled as panels using Fiji (69 (link)) and Adobe Photoshop CC 2021.

For staining of nuclei, parasites were incubated with 1 µg/mL DAPI (Sigma, St. Louis, MO, USA) in culture medium for 15 minutes at 37°C. PI-PLC cKO parasites were imaged using a Nikon Eclipse Ni-E widefield microscope equipped with a Hamamatsu C11440 digital camera and a 100×/1.45 NA oil immersion objective. All other parasite lines were imaged on a Leica D6B fluorescence microscope equipped with a Leica DFC9000 GT camera and a Leica Plan Apochromat 100×/1.4 oil objective. Image processing was performed using ImageJ.

All fluorescence images were captured using a Zeiss Axioskop 2 Plus microscope with a Hamamatsu digital camera (model C4742-95) or a Leica D6B fluorescence microscope equipped with a Leica DFC9000 GT camera and a Leica Plan Apochromat 100×/1.4 oil objective.
Microscopy of live-parasite-infected erythrocytes was performed as previously described (93 (link)). Briefly, parasites were incubated in standard culture medium with 1 μg/mL Hoechst 33342 (Invitrogen) for 15 min at 37°C prior to imaging. A 5.4-μL portion of infected erythrocytes was added on a glass slide and covered with a coverslip. Nuclei were stained with 1 μg/mL Hoechst 33342 (Invitrogen). Microtubules were visualized by incubation of parasites in medium containing 1:1,000 TubulinTracker deep red (Thermo Fisher Scientific; dissolved in dimethyl sulfoxide [DMSO]), which labels polymerized tubulin, for 15 min at 37°C prior to imaging, as previously described (94 ). Images were processed using Fiji (91 (link)), and Adobe Photoshop CC 2021 was used for display purposes only.