Fig. 6

HMH reduces the mitochondrial membrane potential (ΔΨm) depolarization and mtROS production. RAW 264.7 macrophages (1 × 10⁵ cells/mL) were treated with 0–10 mg/mL HMH for 2 h prior to LPS (200 ng/mL) treatment for 24 h. (A) Cells were harvested and stained using a Muse Mitopotential Kit, and the ΔΨm was measured using flow cytometry. (B) Total ΔΨm depolarized cells (%) were shown. (C) In a parallel experiment, cells were stained using 5 μM MitoTracker Green for 30 min and subsequently with 10 μM MitoSOX Red for 10 min in the presence or absence of 5 μM MitoTEMPO. Fluorescence images were captured using a CELENA S Digital Imaging System. (D) The 3 dpf larvae were microinjected with LPS (0.5 mg/mL, 2 nL per larva) and treated with 0–5 mg/mL HMH for 24 h. Larvae were stained using 10 μM MitoSOX Red for 30 min and 5 μM MitoTracker Green for 10 min in embryo medium in the presence or absence of 10 μM MitoTEMPO, and the images were captured using a CELENA S Digital Imaging System. ∗∗∗p < 0.001 vs. untreated cells and ^###p < 0.001 vs. LPS-treated cells.