Fig. 1

Generation of L. major ΔLABCG1-2 parasites and accumulation of NBD-PS. a Restriction map of the native LABCG1-2 locus. Hybridisation probes are shown by arrowheads and codon regions by black boxes. Black arrows indicate the direction of the transcription. Single allele LABCG1-2 mutant parasites were generated using hygromycin replacement construct and the remaining single allele was replaced by homologous recombination. b Southern blot analysis of wild-type (WT) and ΔLABCG1-2 lines. DNA was digested with AgeI and subjected to Southern blot analysis using radiolabelled probes that recognise the intergenic region of LABCG1 and LABCG2 (probe 1, b1) and the 5′ UTR region of LABCG2 (probe 2, b2). The positions of molecular markers (Kbp) are indicated on the right. c Promastigotes of L. major lines: control (WT), ΔLABCG1-2, and add-back parasites with LABCG1, LABCG2 and LABCG1-2 (ΔLABCG1-2 + LABCG1, ΔLABCG1-2 + LABCG2 and ΔLABCG1-2 + LABCG1-2) were incubated with NBD-PS for 30 min at 28 °C. After extraction with BSA and washing with PBS, the intracellular fluorescence was determined by flow cytometry. Data are the mean ± SD of three independent experiments. Statistical differences relative to the control values were calculated using Student’s t-test (*P < 0.05)