Various P. berghei-ANKA lines were used to infect mice and cultured cells. Transgenic lines used in this study included PbHsp70mCherry (referred to as PbmCherry), which constitutively expresses mCherry in the parasite cytosol (Burda et al., 2015); Pb Hsp70mCherry ef1αNLuc (referred to as PbNLuc), which constitutively expresses mCherry and the luminescent protein NanoLuc (De Niz et al., 2016); PbLisp2Exp1-mCherry (Graewe et al., 2011), which expresses mCherry-tagged EXP1 under the liver stage-specific Lisp2 promoter, enabling visualization of the PVM from 36 hpi onwards; and PbUIS4 UIS4-mCherry (Grützke et al., 2014), which expresses the mCherry-tagged PVM protein UIS4 under its endogenous promoter and enables PVM visualization from early times post-infection. We also used the parasite mutants PbΔUIS4 and PbΔUIS3 (Mueller et al., 2005a,b).

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