Fig. 3: Infection with CSP_mut induces hemocytes to release TEP1 in a Toll-dependent manner.

a The recruitment of hemocytes, which were stained with a hemocyte-specific lipophilic dye (Vybrant CM-DiI), to the midgut of mosquitoes at day 7 after infection with Plasmodium yoelii CSP_wt or CSP_mut parasites (left), and statistical analysis of hemocytes recruited to the midguts (n = 64) (right); scale bar 50 μm. b A representative immunofluorescent confocal image of granulocyte, oenocytoid, and prohemocyte from P. yoelii CSP_wt or CSP_mut parasite-infected mosquitoes (n = 60) at day 5 PI, stained with anti-TEP1 antibody (left), the percentage of TEP1 positive cells in granulocyte, oenocytoid, and prohemocyte was quantified (right). Scale bar 2 μm. c The percentage of melanized mature oocysts (left) in mosquitoes (n = 45), the average number of hemolymph sporozoites (middle) in mosquitoes (n = 45), and the expression of TEP1 (right) in mosquitoes (n = 45) infected with P. yoelii CSP_wt and CSP_mut, pretreated with or without clodronate liposome. The mRNA levels of TEP1 in CSP_mut parasite-infected mosquitoes (n = 15) determined using real-time PCR, after GNBP-B1 (d) or Rel1 (e) knockdown. The percentage of melanized mature oocyst in CSP_mut parasite-infected mosquitoes (n = 45) determined after knockdown of GNBP-B1 (f) or Rel1 (g). The number of hemolymph sporozoites in CSP_mut parasite-infected mosquitoes (n = 45) counted after knockdown of GNBP-B1 (h) or Rel1 (i). For box-plot diagram (a): middle line represents median; boxes extend from the 25th to 75th percentiles. The whiskers mark the 10th and 90th percentiles. b, d–f A two-sided Student’s t-test was used for the statistical analysis. a, c, g–i A two-sided Mann–Whitney U test was used for the statistical analysis. The data were presented as the means ± SD; ns, no significance; p-values were shown. The data were combined based on three independent experiments (a–i). Source data are provided as a Source Data file.