ferrozine hy 137805  (MedChemExpress)

Journal: Advanced Science

Article Title: Lysosome‐Featured Cell Aggregate‐Released Extracellular Vesicles Regulate Iron Homeostasis and Alleviate Post‐Irradiation Endothelial Ferroptosis for Mandibular Regeneration

doi: 10.1002/advs.202505070

Figure Lengend Snippet: Lysosomal dynamics is driven by HIF‐2α signaling and regulates iron homeostasis in CAs. A) Immunostaining for HIF‐1α, HIF‐2α, or HIF‐3α (red), with nuclei stained with DAPI (blue). Scale bars, 50 µm. B–D) Quantifications of HIF‐1α, HIF‐2α, and HIF‐3α fluorescent intensity in (A). SC, unaggregated stem cell; CA, cell aggregate. E) The inhibitory efficacy of siRNAs on EPAS1 (encoding HIF‐2α) expression levels in CAs was evaluated by qRT‐PCR. F) Fluorescent staining of AIE‐lysosome (red) in CAs, with nuclei stained with Hoechst (blue). Scale bars, 50 µm. G) Quantification of AIE‐lysosome fluorescent intensity in (F). H) Fluorescent staining of the pH indicator, lysosensor DND‐189 (green) in CAs, with nuclei stained with Hoechst (blue). Scale bars, 50 µm. I) Quantification of DND‐189 fluorescent intensity in (H). siCtrl, siRNA negative control; siEPAS1, siRNA oligonucleotides of endothelial PAS domain‐containing protein 1 ( EPAS1 ). (J) Fluorescent staining of AIE‐lysosome (red), with nuclei stained with Hoechst (blue). Scale bars, 50 µm. K) Quantification of AIE‐lysosome fluorescent intensity in (J). L) Fluorescent staining of DND‐189 (green) in CAs, with nuclei stained with Hoechst (blue). Scale bars, 50 µm. M) Quantification of DND‐189 fluorescent intensity in (L). DMSO, dimethyl sulfoxide; PT2385, a HIF‐2α inhibitor. N) Fluorescent staining of the ferrous iron (Fe 2+ ) indicator, FerroOrange (red), with nuclei stained with Hoechst (blue). Scale bars, 50 µm. O) Quantification of FerroOrange fluorescent intensity in (N). P) Intracellular Fe 2+ content was analyzed by a colorimetric method. BafA1, bafilomycin A1, a lysosomal V‐ATPase inhibitor. Q) Immunostaining for Col1a1 or fibronectin (red), with nuclei stained with DAPI (blue). Scale bars, 50 µm. R,S) Quantifications of Col1a1 and fibronectin fluorescent intensity in (Q). T) ECM stiffness of CAs analyzed by the AFM nanoindentation test. U) Bright‐field images for the process of cell aggregation. Scale bars, 150 µm. V) Quantification of cell aggregation efficiency. Ferrozine, a Fe 2 ⁺ chelator. Results are expressed as mean ± SD. n = 3 samples per group for each experimental readout. p values were calculated using Student's t ‐test (B,C,D,E,G,I,K,M,R,S,T), one‐way ANOVA with Tukey's post hoc test O,P), or two‐way ANOVA (V). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Bafilomycin A1 (BafA1; MedChemExpress, USA; Cat#HY‐100558), GW4869 (MedChemExpress, USA; Cat#HY‐19363), PT2385 (MedChemExpress, USA; Cat#HY‐12867), and ferrozine (MedChemExpress, USA; Cat#HY‐137805) were dissolved in dimethyl sulfoxide (DMSO; Beyotime, China; Cat#C2041S‐2).

Techniques: Immunostaining, Staining, Expressing, Quantitative RT-PCR, Negative Control

Journal: Advanced Science

Article Title: Taurine Inhibits Ferroptosis Mediated by the Crosstalk between Tumor Cells and Tumor‐Associated Macrophages in Prostate Cancer

doi: 10.1002/advs.202303894

Figure Lengend Snippet: Taurine protects PCa against ferroptosis by activating the LXRα/SCD1 axis. A) The antioxidant activity of taurine was analyzed by a DPPH assay. Liproxstatin‐1 was used as a positive control. B) The iron chelator activity of taurine was analyzed by using the ferrozine Fe 2+ binding assay. Deferoxamine (DFO) was used as a positive control. C) The LXRα expression in both cytoplasm and nuclear in PCa cells after TauT knockout. D) Cell viability analysis of PCa cells after LXRα knockout following with the RSL3 treatment for 24 h. E) Cell viability analysis of PCa cells after TauT knockout combined with GW3965 incubation following with the RSL3 treatment for 24 h. F,G) The mRNA (F) and protein expression (G) of SCD1 in PCa cells after LXRα knockout. H) Binding sites of LXRα transcription factors in the promoter region of SCD1. A black triangle indicates binding site(s) for LXRα transcription factors. Experiments presented in (C–H) were performed with the existence of taurine (100 µ m ). Each experiment was performed in triplicate and independently repeated three times. (Two‐tailed Student's t ‐test was used for the statistical analysis: ns, not significant; ** , p < 0.01; *** , p < 0.001. Data are presented as means ± SD, n = 3).

Article Snippet: Then, 3‐(2‐pyridyl)−5,6‐diphenyl‐1,2,4‐triazine‐p, p′‐disulfonic acid monosodium salt hydrate (FerroZine) (MCE, HY‐137805) was added and mixed to achieve a final concentration of 20 µ m . The samples were thoroughly mixed and incubated at room temperature for 1 h. Deferoxamine (DFO) (MCE, HY‐B0988) was used as a positive control.

Techniques: Antioxidant Activity Assay, DPPH Assay, Positive Control, Activity Assay, Binding Assay, Expressing, Knock-Out, Incubation, Two Tailed Test