Scheme of identifying immunoactive compounds of Ganoderma lucidum spores by molecular networking, zebrafish assays and chemometrics.

Assessment of immunoactivity of Ganoderma lucidum spores on zebrafish models of neutropenia. (A) Scheme of zebrafish husbandry and treatment. (B) Representative fluorescent images of Tg (mpx:GFP) transgenic larvae of control and model groups. (C) Neutrophils count in zebrafish of different groups. (D) Neutrophil recovery rate (%) of different groups. BGLS, sporoderm-broken Ganoderma lucidum spores; RGLS, sporoderm-removed Ganoderma lucidum spores. Data are expressed as mean ± SEM, n = 10. *P < 0.05, **P < 0.01 vs. Model.

Assessment of immunoactivity of Ganoderma lucidum spores on zebrafish models of macrophage deficiency and macrophage phagocytosis. (A) Overview of zebrafish development and experimental protocol. (B) Represent images of macrophages in healthy fish and zebrafish model of macrophage deficiency. (C) Macrophages count in zebrafish of different groups. (D) The number of macrophages that phagocytize active carbon nanoparticles of different groups. (E) Macrophage formation efficiency (%) of different groups. (F) Macrophage phagocytosis efficiency (%) of different groups. BGLS, sporoderm-broken Ganoderma lucidum spores; RGLS, sporoderm-removed Ganoderma lucidum spores. Data are expressed as mean ± SEM, n = 10. **P < 0.01, ***P < 0.001 vs. Model.