c11 bodipy lipid peroxidation sensor  (MedChemExpress)

Journal: Communications Biology

Article Title: Hemin-induced transient senescence via DNA damage response: a neuroprotective mechanism against ferroptosis in intracerebral hemorrhage

doi: 10.1038/s42003-025-07983-3

Figure Lengend Snippet: a Western blot analysis was conducted with or without p21i to examine the expression of p21, p53, HO-1, and cleaved PARP proteins at the specified time points after hemin (10 µM) treatment. The quantitation data presented as mean ± s.e.m. from three independent experiments. b Western blot analysis was performed with or without HO-1 knockdown to examine the expression of HO-1, p53, and cleaved caspase-3 proteins at the indicated time points after hemin treatment. The quantitation data presented as mean ± s.e.m. from three independent experiments. c Western blot analysis was carried out after transfecting with or without HO-1 expression plasmid to examine the expression of HO-1, p21, and GPX-4 proteins at the indicated time points after hemin treatment. The quantitation data presented as mean ± s.e.m. from three independent experiments. d Measuring cellular lipid peroxidation in SH-SY5Y cells transfected with or without the HO-1 plasmid and treated with or without hemin using the C11 BODIPY™ 581/591 fluorescent probe. Total C11 BODIPY™ 581/591 (red), oxidized C11 BODIPY™ 581/591 (green), and Hoechst-stained nuclei (blue). Increased green fluorescence suggests higher lipid peroxidation, while the red fluorescence reflects areas of non-oxidized lipid content. Scale bar = 20 µm. Zoomed image is shown at 3x magnification. e Viability assay using CellTiter-Glo reagent showing results for control cells, HO-1 knockdown cells, and HO-1 overexpressing cells at indicated time points after hemin treatment. The quantitation data presented as mean ± s.e.m. from three independent experiments. f A schematic illustration of the sequential cellular responses to hemin exposure and its significance in counteracting ICH-associated hemin and iron toxicity. Any interference with these sequential processes, such as inhibition of transient senescence or altering the timing of HO-1 induction without corresponding co-expression with senescence, can lead to unsynchronized cellular response and cause ferroptotic and/or apoptotic cell death. All statistical analyses were performed by two-sided Student’s t-test, p-values are indicated in the respective graph.

Article Snippet: The BODIPY 581/591 C11 Lipid Peroxidation Sensor assay (Cell Signaling Technology, # 95978) was conducted following the manufacturer’s instructions.

Techniques: Western Blot, Expressing, Quantitation Assay, Knockdown, Plasmid Preparation, Transfection, Staining, Fluorescence, Viability Assay, Control, Inhibition

Journal: Communications Biology

Article Title: Hemin-induced transient senescence via DNA damage response: a neuroprotective mechanism against ferroptosis in intracerebral hemorrhage

doi: 10.1038/s42003-025-07983-3

Figure Lengend Snippet: iPSC-derived neurons were exposed to PEG-OAC and DEF-OAC-PEG nanoparticles 1 h after hemin (10 μM) treatment. a Representative images and quantification of SA-β-Gal positive cells per field in iPSCs-derived neurons treated with or without hemin and nanoparticles (PEG-OAC and DEF-OAC-PEG) for 24 h, Scale bar = 20 µm. The data presented is derived from three independent experiments. b Viability assay (CellTiter-Glo) of iPSC-derived neurons treated with or without hemin and nanoparticles (PEG-OAC and DEF-OAC-PEG) for 24 h, the data presented is derived from three independent experiments. c Western blot analysis to assess the expression of p-ATM (S1981), HO-1, p21, cleaved PARP, and GPX-4 proteins at indicated time points, the data presented is derived from three independent experiments. d Representative image of the lipid peroxidation assay using the C11 BODIPY™ 581/591 fluorescent probe in iPSC-derived neurons following 24-hour treatment with or without hemin and nanoparticles (PEG-OAC and DEF-OAC-PEG). Total C11 BODIPY™ 581/591 (red), oxidized C11 BODIPY™ 581/591 (green), and Hoechst-stained nuclei (blue). Increased green fluorescence suggests higher lipid peroxidation, while the red fluorescence reflects areas of non-oxidized lipid content. Scale bar = 20 µm. Zoomed image is shown at 3x magnification. All statistical analyses were performed by one-way ANOVA, p-values are indicated in the respective graphs.

Article Snippet: The BODIPY 581/591 C11 Lipid Peroxidation Sensor assay (Cell Signaling Technology, # 95978) was conducted following the manufacturer’s instructions.

Techniques: Derivative Assay, Viability Assay, Western Blot, Expressing, Peroxidation Assay, Staining, Fluorescence