Characterization of <i>Plasmodium pp1</i> (<i>PBANKA_102830</i>).

<p><i>(A) pp1</i> mRNA levels during the <i>P</i>. <i>berghei</i> life cycle. <i>Pbpp1</i> mRNA levels were analyzed by real-time PCR using cDNAs from the different stages of <i>P</i>. <i>berghei</i>. The <i>arginyl-tRNA synthetase</i> (<i>PbArgRS</i>, <i>PB000094</i>.<i>03</i>.<i>0</i>) was used as internal control. Each value is the mean ± SD of two independent experiments. MG: midgut; SG: salivary gland; Liver: liver stages; BS: asexual blood stages; Gtc: gametocyte. <i>(B) Pbpp1</i> mRNA levels in <i>Pbpp1</i> cKO Ssp were quantified by qPCR. P = 0.041. P value was calculated by t test. Shown are mean ± SD of two independent experiments. <i>(C)</i> Immunoblot analysis of <i>Pbpp1</i> cKO and wt TRAP/FlpL(-) Ssp using anti-PbPP1 serum. CSP was used as control. <i>(D)</i> The liver stage development of <i>Pbpp1</i> cKO Ssp in HepG2 cells was indistinguishable from the wild type TRAP/FlpL(-) Ssp. Hepatic parasites were stained with anti-PbHSP70 and anti-PbPP1 antibodies at 6h, 24h, 36h, 48h, and 55h post-infection. Bar, 10 μm. <i>(E)</i> eIF2α phosphorylation level in <i>Pbpp1</i> cKO sporozoites was indistinguishable from wild type. Five hundred thousand wt TRAP/FlpL(-) or <i>Pbpp1</i> cKO sporozoites were dissected from mosquito salivary glands. Levels of PbeIF2α-P and total PbeIF2α were quantified by densitometry analysis of immunoblots performed with antibodies against anti-eIF2α-P and anti-total eIF2α [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005370#ppat.1005370.ref015" target="_blank">15</a>, <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1005370#ppat.1005370.ref016" target="_blank">16</a>]. Values are shown below the bands. Results were similar in two independent experiments.