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Fig. S1. Micrographs of P. berghei wild-type (WT) and LipB-deficient (PbΔLipB) asexual blood stage parasites labelled with antibodies to the apicoplast marker ACP and stained with the nuclear dye DAPI. Parasites were harvested from infected mice and cultured overnight prior to fixation, staining and imaging (see Experimental procedures). WT and PbΔLipB parasites showed no evident differences in asexual blood stage morphology, including the apicoplast organelle. These data are provided as an extension of Fig. 2C. Space bar, 10 μm.

Fig. S2. Images of P. berghei wild-type (WT) and LipB-deficient (PbΔLipB) liver stage parasites sampled at 48 h post invasion. Parasites were stained with the nuclear dye DAPI, the MitoTracker Red dye, and anti-lipoic acid (LA) antibodies to determine lipoylated structures. WT and PbΔLipB parasites were treated with DMSO or the lipoic acid analogue 8-BOA. PbΔLipB parasites showed little to no remaining lipoylated structures in the parasite after 8-BOA treatment. Each row represents different stainings for a given parasite. Images are provided as an extension of Fig. 4. Scale bar, 10 μm.

Fig. S3. Antibody staining of P. berghei wild-type (WT) and LipB-deficient (PbΔLipB) liver stage parasites sampled at 48 h post invasion. Parasites were stained with the nuclear dye DAPI and antibodies to lipoic acid (LA) to visualize lipoylated proteins within the parasite and the host. The apicoplast was labelled with antibodies to the apicoplast marker Acyl-Carrier Protein (ACP). PbΔLipB parasites showed less lipoylation in the apicoplast as compared to WT parasites (see greater extent of orange overlays in WT parasites compared to more green staining in the LipB-deficient parasites). Pertains to Fig. 4. Space bar, 10 μm.

Fig. S4. Images of P. berghei wild-type (WT) and LipB-deficient (PbΔLipB) liver stage parasites sampled at 48 h post invasion (hpi). Parasites were stained with the nuclear dye DAPI and anti-EXP-1 antibodies to determine the morphology of the parasitorphorous vacuole. Anti-ACP antibodies were used to determine the morphology of the apicoplast organelle. PbΔLipB parasites showed smaller and less branched apicoplasts during liver stage development. Each column contains images from a single parasite. Data are provided as an extension of Fig. 5. Space bar, 10 μm.

Fig. S5. Images of P. berghei wild-type (WT) and LipB-deficient (PbΔLipB) liver stage parasites sampled at 54 h post invasion (hpi). Parasites were stained with the nuclear dye DAPI and anti-MSP-1 antibodies to determine the morphology of the parasitorphorous vacuole. Anti-ACP antibodies were used to determine the morphology of the apicoplast organelle. PbΔLipB parasites showed smaller and less branched apicoplasts during liver stage development. Data are provided as an extension of Fig. 5. Space bar, 10 μm.

Fig. S6. Images of P. berghei wild-type (WT) and LipB-deficient (PbΔLipB) liver stage parasites during late schizont development sampled at 54 h post invasion. Parasites were stained with the nuclear dye DAPI and anti-MSP-1 antibodies to define the parasite plasma membrane. PbΔLipB parasites showed major defects in plasma membrane morphology. Data are provided as an extension of Fig. 6C. Space bar, 10 μm.

Fig. S7. Images of P. berghei wild-type (WT) and LipB-deficient (PbΔLipB) liver stage parasites sampled at 62 h post invasion. Parasites were stained with the DAPI nuclear dye and anti-MSP-1 antibodies to define the parasite plasma membrane. PbΔLipB parasites showed major defects in plasma membrane morphology. Data are provided as an extension of Fig. 6C. Scale bar, 10 μm.

Table S1. List of oligonucleotides used in this study.

Table S2. Mean percent reticulocytes in clofibrate- or mocktreated mice.

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