Various P. berghei ANKA lines were used to infect mice. Lines used in this study included Pb mCherryHsp70, constitutively expressing mCherry in the parasite cytosol (34); a transgenic line expressing NanoLuc luciferase and mCherry under constitutive promoters (Pb mCherryHsp70NLucef1α) (35); a transgenic line expressing firefly luciferase and mCherry under constitutive promoters Pb mCherryHsp70FLucef1α (36); and the nonsequestering P. berghei line PbΔSBP1-GFP-mCherryHsp70FLucef1α (9). In addition, P. berghei lines G623 (820) and G629 (GNPm9), with CFP expression under the PBANKA_101870 promoter, and lines G488 (820), G458 (GNPm9), and G455(HP), with CFP expression under the Hsp70 promoter were used to investigate differences in parasite localization among previously published gametocyte-producing and gametocyte-nonproducing lines (15). Lines G629 (GNPm9) and G455(HP) were used for normalization purposes. Subclones of the above lines were generated by detached cell injection, as previously described (37).

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