Fig. 1

The nuclear PfArp4 is an essential protein in P. falciparum.a Schematic representation of generation of transgenic PfArp4-Ty1-Ribo line. Co-transfection of the plasmid pUF1-BSD-cas9 with pL6CS-hDHFR-PfArp4 leads to integration of the PfArp4-Ty1-Ribo containing cassette in the endogenous locus. b PCR validation of PfArp4-Ty1-Ribo lines. The sequences of primer pairs are listed in Additional file 10: Table S4. c Growth curve assay of wild-type 3D7-G7 parasites with or without GlcN in the culture (n = 3, bars are SD). d Growth curve assay of PfArp4-Ty1-Ribo (7A clone) parasites with or without GlcN in the culture (n = 3, bars are SD). e Western blot of protein extracts from ring, trophozoite and schizont stage PfArp4-Ty1-Ribo parasites with (+) or without (−) GlcN in the culture for 2 cycles. f Representative Giemsa-stained blood smears of ring (R, 8–12 h), trophozoite (T, 28–32 h) and schizont (S, 40–44 h) stagePfArp4-Ty1-Ribo lines with (+) or without (−) GlcN in the culture for 3 cycles. g Schematic representation of the dual-tagging transgenic parasite line, PfArp4-Ty1::PfArp6-HA. h Co-IFA analysis of PfArp4-Ty1::PfArp6-HA line at ring, trophozoite, or schizont, respectively. i Co-IP analysis of PfArp4-Ty1::PfArp6-HA line and 3D7_G7 line (negative control) with anti-PfAldolase, anti-HA and anti-Ty1 antibodies for IP. Abbreviations: BSD, blasticidin-S deaminase; sgRNA, a synthetic single-guide RNA; hDHFR, human dihydrofolate reductase; RHL, 3′ flanking fragment for crossover recombination; LHL, 5′ flanking fragment for crossover recombination; Tag, insertion sequence. Scale-bars: f, 2 μm; h, 1 μm