Journal: Allergologia et immunopathologia

Article Title: Alpinia officinarum Hance extract relieved sepsis-induced myocardial ferroptosis and inflammation by inhibiting lncRNA MIAT/TRAF6/NF-κB axis.

doi: 10.15586/aei.v52i5.1035

Figure Lengend Snippet: Figure 2 Alpinia officinarum Hance extract inhibited sepsis-induced myocardial ferroptosis. Groups were divided into the Sham, LPS, LPS+25 mg/kg AOE, LPS+50 mg/kg AOE, and LPS+100 mg/kg AOE groups. (A) The Fe2+ level was evaluated through the corresponding commercial kit. (B) The levels of MDA and SOD were examined through ELISA. (C) The protein expressions of ACSL4 and GPX4 were tested through western blot.

Article Snippet: Detection of Fe2+ The Fe2+ assay kit (E-BC-K304-S, Elabsciences, China) was used for measuring Fe2+ level as per manufacturer’s protocols.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Allergologia et immunopathologia

Article Title: Alpinia officinarum Hance extract relieved sepsis-induced myocardial ferroptosis and inflammation by inhibiting lncRNA MIAT/TRAF6/NF-κB axis.

doi: 10.15586/aei.v52i5.1035

Figure Lengend Snippet: Figure 5 Overexpression of MIAT reduced the cardioprotective effect of Alpinia officinarum Hance extract. Groups were separated into the Sham, LPS, LPS+100 mg/kg AOE, and LPS+100 mg/kg+Ad-MIAT groups. (A) The protein expressions of TRAF6, p-P65, and P65 were verified through western blot. (B) The level of CK-MB in serum was examined through ELISA. (C) The level of cTnl in serum was measured through ELISA. (D) The Fe2+ level was tested through the corresponding commercial kit. (E) The levels of MDA and SOD were measured through ELISA.

Article Snippet: Detection of Fe2+ The Fe2+ assay kit (E-BC-K304-S, Elabsciences, China) was used for measuring Fe2+ level as per manufacturer’s protocols.

Techniques: Over Expression, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Viruses

Article Title: Infectious Spleen and Kidney Necrosis Virus Triggers Ferroptosis in CPB Cells to Enhance Virus Replication.

doi: 10.3390/v17050713

Figure Lengend Snippet: Figure 3. ISKNV infection induces ferroptosis in CPB cells. (A) Transmission electron microscopy of CPB cells treated with DMSO (72 h), erastin (10 µmol/L, 72 h), and ISKNV (100 MOI, 24 h, 48 h, 72 h). Scale bars = 1 µm. (B) Analysis of Fe2+ levels in CPB cells after treatment with ISKNV (100 MOI) using the fluorescent probe FerroOrange by laser scanning confocal microscopy. Scale bars = 20 µm. (C) Quantitative analysis of the mean fluorescence intensity of (B) using Image J. (D) Analysis of intracellular ROS levels using DCFH-DA staining, and laser scanning confocal microscopy of CPB cells treated with ISKNV (100 MOI) for 24–72 h. Scale bars = 10 µm. (E) Quantitative analysis of the mean fluorescence intensity of (D) using Image J. (F–H) Detection of Fe2+, ROS, and MDA levels in cell lysates treated with ISKNV (100 MOI) for 24–72 h by microplate reader. * p < 0.05, ** p < 0.01, and *** p < 0.001, with p > 0.05 considered not significant (ns).

Article Snippet: The Fe2+ content of the cells was detected by laser scanning confocal microscopy and a microplate reader using the fluorescent probe FerroOrange (Elabscience, E-BC-F101, Wuhan, China).

Techniques: Infection, Transmission Assay, Electron Microscopy, Confocal Microscopy, Fluorescence, Staining

Journal: Pharmaceutical Biology

Article Title: Indirubin regulates M1/M2 polarization and inhibits ferroptosis in dextran sulfate sodium induced colitis and in cultured THP-1 cells

doi: 10.1080/13880209.2025.2568215

Figure Lengend Snippet: IDR inhibited ferroptosis in THP-1 mocytes and in THP-1 derived macrophages with M1- or M2- polarized status. (a–d) IDR inhibited RSL3 induced cell loss in THP-1 derived macrophages, including M0 (a), M1 (b), and M2 (c) polarized macrophages, as well as in THP-1 monocytes (d). The viable cell counts were determined with CCK-8 assay. Data were presented as Mean ± S.D. (n = 4). * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to the RSL3 group. (e) The representative plots of IDR inhibiting RSL3 induced cell death determined by PI staining with flow-cytometry analysis. The proportion of PI-positive cells (f) and the preserved cell counts normalized to count beads (g) were also shown in the histogram plots. Data were presented as Mean ± S.D. (n = 3). * p < 0.05, ** p < 0.01. (h) Reactive oxygen species (ROS) levels were determined with DHCF-DA. Data were presented as Mean ± S.D. (n = 3). * p < 0.05, ** p < 0.01. (i) Malondialdehyde (MDA) levels were determined. Data were presented as Mean ± S.D. (n = 3). * p < 0.05, ** p < 0.01. (j) Ferrous iron (Fe 2+ ) levels were quantified using a fluorometric assay. Data were normalized to cell counts and presented as Mean ± S.D. (n = 3). # p < 0.05 compared to the NC group. *** p < 0.05 compared to the DSS group. (k) The effects of IDR treatments on the mRNA expression levels of ferroptosis-associated genes, including Gpx4, Acsl4, Nrf2, and Slc7a11. Data were normalized to the level of reference gene Gapdh and presented as Mean ± S.D. (n = 3). * p < 0.05, ** p < 0.01 compared to the RSL3 group. (l) The representative images of Western blotting analysis of ferroptosis-associated proteins, including 4-HNE, GPX4, NRF2, FTL, and GAPDH. (m) Column plots of 4-HNE, GPX4, NRF2 and FTL abundance after normalizing to GAPDH. Mean ± S.D. (n = 3). * p < 0.05 compared to the RSL3 group.

Article Snippet: A Cell Ferrous Iron (Fe2+) Assay Kit (Fluorometric) (HY-K0322, MedChemExpress LLC, China) was applied to quantify iron concentration in THP-1 cells according to the manufacturer’s manual.

Techniques: Derivative Assay, CCK-8 Assay, Staining, Flow Cytometry, Expressing, Western Blot

Journal: Pharmaceutical Biology

Article Title: Indirubin regulates M1/M2 polarization and inhibits ferroptosis in dextran sulfate sodium induced colitis and in cultured THP-1 cells

doi: 10.1080/13880209.2025.2568215

Figure Lengend Snippet: IDR augmented M2-polarization and inhibited ferroptosis in peritoneal macrophages. (a) The effects of IDR treatments on the surface staining of Arg-1 in M2-polarized mouse peritoneal macrophages were determined by flow-cytometry analysis. (b) Histogram plot of the proportion and MFI (mean fluorescence intensity) of Arg-1 positive cells. Data were presented a Mean ± S.D. (n = 3). ## p < 0.01 compared to the NC group. * p < 0.05 and ** p < 0.01 compared to the M2 group. (c) The viable cell counts were determined with CCK-8 assay. Data were presented as Mean ± S.D. (n = 5). ## p < 0.01 compared to the NC group. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to the M2 group. (d) The representative images of cellular ferrous iron fluorescence. (e) Histogram plot of the MFI of ferrous iron. Data were presented as Mean ± S.D. (n = 5). ### p < 0.01 compared to the NC group. ** p < 0.01 and *** p < 0.001 compared to the RSL3 group.

Article Snippet: A Cell Ferrous Iron (Fe2+) Assay Kit (Fluorometric) (HY-K0322, MedChemExpress LLC, China) was applied to quantify iron concentration in THP-1 cells according to the manufacturer’s manual.

Techniques: Staining, Flow Cytometry, Fluorescence, CCK-8 Assay