Fig 2: Effect of griseofulvin treatment on CM pathogenesis.a Mortality curves of PbWT-infected mice treated with different dosages of griseofulvin (***P < 0.001, log-rank (Mantel–Cox) test). b Growth curve analysis (n = 12) for 2 mg dose per day on day 4, 5, 6, 7, and 8 (mean ± SD; n.s not significant, Two-way ANOVA). c Phosphorimager and scanned images of TLC performed for 14C-ALA labeled parasite free heme and the radioactive counts measured for three different experiments (mean ± SD; n.s not significant, *P < 0.05, unpaired t-test; two-sided). Individual data points are shown as black circles. Full-length scans are provided in the Supplementary Fig. 7. d Extravasation of Evans blue and its quantification (n = 3) (mean ± SD; **P < 0.01, unpaired t-test; two-sided). Individual data points are shown as black circles. e H&E staining of the brain sections. Images were captured using 10x objective. Scale bar = 50 µm. n = 3 independent experiments. f Parasite accumulation g CD3+ cells in the blood vessels. n = 2 independent experiments. h Axonal injury in the brain sections. Images were captured using 20x objective. Scale bar = 20 µm. n = 3 independent experiments. i Giemsa-stained parasites. Images were captured using 100x objective. Scale bar = 5 µm. n = 3 independent experiments. j Hz content in differential interference contrast (DIC; left) and bright field images (right) of paraformaldehyde-fixed RBCs. Images were captured using 100x objective. Scale bar = 5 µm. n = 3 independent experiments. k Parasite Hz (n = 8 for control; n = 5 for treated). l Parasite free heme (n = 5). m Plasma free heme (n = 6). n Plasma hemopexin (n = 6). o Plasma heme/hemopexin ratio. p Plasma hemoglobin (n = 6). k–p (mean ± SD; n.s not significant, **P = 0.01, ***P < 0.001, unpaired t-test; two-sided). q Mortality curves for PbWT-infected C57BL/6 female mice treated with a,ß-arteether alone (n = 9) and a,ß-arteether in combination with griseofulvin (n = 9). The data represent two different batches. (***P < 0.001, n.s not significant, log-rank (Mantel–Cox) test). Source data are provided as a Source Data file.

Fig 3: Culturing T cells and BMDMs(A) Differentiated, confluent BMDMs 6 days after culturing the initial bone marrow-dervied cells.(B) Representative image of cultured splenic T cells in an anti CD3/CD28-coated U-bottom well.(C) Differentiated BMDMs before being primed with 100 ng/mL LPS.(D) Differentiated BMDMs after the first 2 h LPS priming. Loss of ramifications and the swelling of the cell body indicates successful priming (white arrowheads, shown before in e.g., Kawakami et al., 2016). In panel (A), (C) and (D) scale bars represent 100 µm. In panel B scale bar represents 1 mm.

Fig 4: IL-31R signaling suppressed the proliferation of CD4+ T cells. Purified CD4+ T cells from WT and IL-31RA KO mice were stimulated with anti-CD3/anti-CD28. The activation (n=4) (A) and proliferation (n=4) (B) of CD4+ T cells were assayed by flow cytometry. All values are plotted and lines indicate means. Statistical significance was determined by two-tailed Student's t- tests. The experiment was repeated twice. Ctrl, control; Act, activation.

Fig 5: E-sel/AAV2/2 modulates inflammatory gene profile in ischemic muscle without inducing significant inflammatory cell recruitment. (A) PCR array from muscle harvested on POD 21 demonstrating an overall downregulation of inflammatory genes by E-sel/AAV2/2 gene therapy. (B) Representative immunofluorescence images of ischemic muscle stained for CD3, a marker of T cells (pointed arrowheads), and (C) Mac-2/Galectin-3, a marker of macrophages (pointed arrowheads). (D) Counting of CD3+ and (E) Mac-2+ cells demonstrating no significant difference in T cell or macrophage recruitment to ischemic muscle treated with E-sel/AAV2/2 and LacZ/AAV2/2. Scale bars represent 50 µm. Data are presented as mean ± SEM and ns, not significant (P > 0.05).