Effect of DMF and its derivatives on the viability and nitric oxide (NO) output in the BV‐2 microglia cell model. (A) Cell viability examined by the CCK‐8 assay, showing identical cytotoxic profiles of DMF and IDMF, and lack of cytotoxicity up to 100 μm for DRF (*P < 0.05 Diroximel vs. IDMF and DMF). (B–D) IDMF (B) inhibits the production of nitric oxide (NO) significantly better than DRF (D) at 2.5 μm and higher doses (P < 0.05). Furthermore, IDMF inhibits the production of NO significantly better than DMF (C) at 5 μm and up. ##P < 0.05 LPS‐stimulated vs. non‐stimulated controls; **P < 0.05 vs. LPS‐stimulated control. Error bars indicate standard deviations. N value: 3. P values were calculated using two‐tailed t‐test.

DMF and its derivatives inhibit the expression of iNOS, COX‐2, TNF‐α, IL‐1β and IL‐6 in LPS‐stimulated BV‐2 cells unequally, as determined by RT‐PCR. *P < 0.05 IDMF vs. DMF and Diroximel; #P < 0.05 IDMF and DMF vs. Diroximel; **P < 0.05 IDMF vs. DMF and Diroximel, and DMF vs. Diroximel. Error bars indicate standard deviations. N value: 3. P values were calculated using two‐tailed t‐test.

Effect of DMF and its derivatives on thymic T cell maturation in the lck‐GFP transgenic zebrafish model. (A) GFP‐labeled T cells in 8‐day‐old lck‐GFP zebrafish are ablated in response to DMF and its derivatives. Arrowheads denote GFP‐labeled cells in the thymus. (B) The GFP fluorescent area in the thymus of lck‐GFP zebrafish. (C) The integrated density of the GFP fluorescence. *P < 0.05 vs. untreated control; **P < 0.01 vs. untreated control. Error bars indicate standard deviations. Scale bar = 250 nm; N value: 15. P values were calculated using two‐tailed t‐test.

Microphotographic documentation of Jurkat cells cultured without (A) or with (B) IDMF (25 μg·mL⁻¹). Initial magnification: ×4, bar: 750 μm. N value: 3.