Journal: iScience

Article Title: HMOX1 drives dihydroartemisinin-sensitized ferroptosis antagonized by mitochondrial fusion

doi: 10.1016/j.isci.2025.114382

Figure Lengend Snippet: Mitochondrial oxidative stress induced by DHA and RSL-3 activates mitochondrial fusion (A) The thumbnail sketch of mitochondrial functions that may be regulated by mitochondrial oxidation. (B) Cell viability of N27 cells treated with DHA (1.5 μM) and RSL-3 (100 nM) for 48 h in the absence or presence of mitochondrial regulators (2 μM oligo A, 2 μM CCCP, 10 μM αKG, 1 μM rotenone), n = 6 wells from one representative of two independent experiments. (C–E) Western blot and quantifications of the OXPHOS, Tom20, β-actin, and GAPDH expression in N27 cells treated with DHA (1.5 μM) and RSL-3 (100 nM) for 12 h. (F–I) Western blot and quantifications of the MFN1, MFN2, DRP1, and GAPDH expression in N27 cells treated with DHA (1.5 μM) and RSL-3 (100 nM) for 12 h. (J) N27 cells were treated with DHA (1.5 μM) and RSL-3 (100 nM) for 12 h and detected by JC-1 using flow cytometry. Statistical analysis of the ratio of the MFI of JC-1 red to JC-1 green is shown, n = 6 wells from one representative of two independent experiments. (K–M) Western blot and quantifications of the MFN1, MFN2, and GAPDH expression in N27 cells treated with MitoQ (5 μM), DHA (1.5 μM), and RSL-3 (100 nM) for 12 h. (N) Cell viability of N27 cells treated with DHA (1.5 μM) and RSL-3 (100 nM) in the absence or presence of mitochondrial fusion promoter M1 (5 μM) for 48 h, n = 6 wells from one representative of two independent experiments. Data are means ± SEM, n = 3 wells from one representative of two independent experiments unless specified. One-way ANOVA was performed unless specified.

Article Snippet: DRP1 (C-terminal) Polyclonal antibody (1:5000) , Proteintech , 12957-1-AP; RRID: AB_2093525.

Techniques: Western Blot, Expressing, Flow Cytometry

Journal: Frontiers in Psychiatry

Article Title: SENP3/FIS1-regulated PFC neural mitochondrial fragmentation underlies the mechanism of electroacupuncture attenuating depressive behavior in CUMS mice

doi: 10.3389/fpsyt.2025.1645757

Figure Lengend Snippet: EA reduces FIS1-regulated PFC neural mitochondrial fragmentation in depressive mice. (A) Representative TEM images of PFC neural mitochondria ultrastructure, Bar = 5μm, 2μm (enlarged), red: neural mitochondria, yellow: autolysosome; (B) Average mitochondrial diameter in PFC neurons in TEM; (C) Average mitochondrial size in PFC neurons in TEM; (D) Histogram of the relative expression of DRP1; (E) Histogram of the relative expression of MFF; (F) Histogram of the relative expression of FIS1; (G) Western blotting images of PFC DRP1, MFF, and FIS1; *P<0.05, ns, no significance.

Article Snippet: After blocking with BSA/TBST at room temperature for 3 h, the membranes were incubated with primary antibodies, including DRP1 (1:1500, 12957-1-Ap, Proteintech, Chicago, USA), MFF (1:4000, 66527-1-Ig, Proteintech), FIS1 (1:1000, SAB2702049, Sigma-Aldrich), SUMO2/3 (1:1500, 11251-1-Ap, Proteintech), SENP3 (1:1000, 17659-1-Ap, Proteintech), and GAPDH (1:3000, HRP-60004, Proteintech), overnight at 4°C.

Techniques: Expressing, Western Blot