MOESM1 of Application of the automated haematology analyzer XN-30 for discovery and development of anti-malarial drugs
Creators: name: Tougan, Takahiro
name: Toya, Yuji
name: Uchihashi, Kinya
name: Horii, Toshihiro
Publisher: name: figshare
Year: 2019
Subjects: subject: Biochemistry
subject: Microbiology
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Cell Biology
subject: Pharmacology
subject: 39999 Chemical Sciences not elsewhere classified
subject: FOS: Chemical sciences
subject: FOS: Chemical sciences
subject: 110309 Infectious Diseases
subject: FOS: Health sciences
subject: FOS: Health sciences
subject: 60506 Virology

description: Additional file 1: Fig. S1. Representative M scattergram of in vitro cultured sample. The horizontal and vertical axes indicate intensities of side fluorescent light (SFL, which corresponds to DNA content) and forward scattered light (FSC, indicating size of iRBCs), respectively. The colours indicate the following: red, ring-form; orange, trophozoite; purple, schizont; and blue, polychromatic RBC. The colours were assigned based on the default setting of the XN-30 analyzer. The scattergram was cited from Fig. 1a(i), DMSO). Fig. S2. Validation in the assay system. (a and b) 24 h, (c and d) 48 h. (a and c) The scatter-plot of the growth inhibition rate. The growth inhibition rate was calculated based on SCHZ-RBC% at 24 h and MI-RBC% at 48 h (see also Methods). The colours indicate the following: blue, 0.5% DMSO; dark red, positive control (5 µM artemisinin); and dark blue, negative control (saline). (b(i) and d(i)) The growth inhibition rate. (b(ii) and d(ii)) The values of validation indices. Abbreviations are as follows: CV %, coefficient of variation; S/B, signal-to-background ratio; and S/N, signal-to-noise ratio. Fig. S3. M scattergrams of the effective compounds, related to Figs. 3 and 4 and Table 1. (a) Type I, (b) Type II, (c) Type III, (d) Type IV. The left and right panels indicate scattergrams at 24 and 48 h, respectively. *, **, and, ** represent effective compounds described in Tong et al. [19] and Dennis et al. [20], and the reference compound mefloquine, respectively. The colours indicate the following: red, ring-form; orange, trophozoite; purple, schizont; and blue, polychromatic RBC. The colours were assigned based on the default setting of the XN-30 analyzer; however, these may be misclassified after compound treatment as described in the Discussion. Fig. S4. Microscopic images of parasites treated with anti-malarial drugs, related to Fig. 1. (a) ART, (b) CQ, (c) DMSO. Sixteen representative images were randomly selected. Scale bar represents 5 µm. Fig. S5. Two possibilities for the generation of a Type II outcome. Possibility 1: The test compound competed with the DNA staining dye in Fluorocell M, as insufficiently stained DNA are likely to show low DNA content. Possibility 2: The test compound fragmented genomic DNA and the DNA was flowed out; the efflux of fragmented DNA reduced the DNA content. Fig. S6. Microscopic images of Type IV outcome, related to Figs. 3 and 4 and Table 1. Typical microscopic images of parasites treated with MMV026550, MMV011765, MMV024443, or MMV030734 after 48 h of incubation. Scale bar represents 5 µm.