Journal: Science signaling

Article Title: ALOX5 drives the pyroptosis of CD4 + T cells and tissue inflammation in rheumatoid arthritis.

doi: 10.1126/scisignal.adh1178

Figure Lengend Snippet: Fig. 2. ALOX5 expression is increased in RA CD4+ T cells. (A) Heatmap shows the expression of RA-related genes in peripheral blood CD4+ T cells from NCs and patients with RA. n = 8 NCs and 10 patients with RA. ALOX5 is highlighted with an arrow and red box. (B) Enrichment analysis for the top 10 diseases and functions represented by the transcripts showing differential expression in the NC and RA groups. Those in which ALOX5 was involved are outlined in red boxes. (C) ALOX5 transcript amounts in NC and RA CD4+ T cells as measured by RT-qPCR. n = 5 patients per group. (D) Representative immunoblot shows ALOX5 abundance in isolated CD4+ T cells from NCs and patients with RA. n = 5 patients per group. β-Actin is a loading control. (E) Quantification of ALOX5 protein in (D). Each protein quantification is based on data combined from three inde- pendent Western blots. n = 5 patients per group. (F) Representative immunoblot shows ALOX5 abundance in CD4+ T cells isolated from patients with RA treated with cortico- steroids (CS+, n = 3 patients) or untreated (CS−, n = 3 patients). ALOX5 protein abundance was quantified on the basis of data from three independent experiments. All data are presented as the mean ± SEM. Statistical significance was determined by Student’s t test; **P < 0.01.

Article Snippet: To explore the potential of ALOX5 pharmacologic inhibition in suppressing T cell pyroptosis, cells were treated with zileuton (MCE, HY- 14164) at 50 μM for 24 hours.

Techniques: Expressing, Quantitative Proteomics, Quantitative RT-PCR, Western Blot, Isolation, Control

Journal: Science signaling

Article Title: ALOX5 drives the pyroptosis of CD4 + T cells and tissue inflammation in rheumatoid arthritis.

doi: 10.1126/scisignal.adh1178

Figure Lengend Snippet: Fig. 7. Association of ALOX5 with clinical remission in RA. (A) Immunoblotting for and quantification of ALOX5 abundance in NCs and patients with RA in dif- ferent phases of disease. β-Actin is a loading control. Each lane represents a single patient. Images are representative of n = 3 independent experiments with samples from different sets of patients. (B) Immunofluorescence staining for ALOX5 and CD3 in human synovial tissue from patients with high disease activity or in remission and quantification of the percentage of CD3+ cells that were also ALOX5+. n = 4 patients per group. (C) Calculated ALOX5 scores for 56 patients with RA with high disease activity or in remission. (D) ROC curve analyses were performed to assess the associations of high RA activity with several clinical parameters. The area under the curve values were 0.9572 for CRP (P < 0.0001), 0.8863 for the ESR (P < 0.0001), 0.7893 for the ALOX5 score (P = 0.0002), 0.6845 for RF (P = 0.0178), and 0.6501 for CCP (P = 0.0540). (E) Patients with RA in the remission and high disease activity groups were stratified into ALOX5high and ALOX5low groups (χ2, P = 0.0002). (F) Correlation matrix of the ALOX5 score and other clinical parameters in high-activity patients and remission patients. The power of the correlation is represented by circle color and size. For the patient data in (C) to (F), n = 27 patients in remission and 29 patients with high disease activity. All data are shown as the mean ± SEM. Statistical significances were determined by one-way ANOVA followed by Bonferroni’s multiple-comparisons test for multigroup comparisons or Student’s t test; *P < 0.05, **P < 0.01, and ****P < 0.0001.

Article Snippet: To explore the potential of ALOX5 pharmacologic inhibition in suppressing T cell pyroptosis, cells were treated with zileuton (MCE, HY- 14164) at 50 μM for 24 hours.

Techniques: Western Blot, Control, Immunofluorescence, Staining, Activity Assay

Journal: Science signaling

Article Title: ALOX5 drives the pyroptosis of CD4 + T cells and tissue inflammation in rheumatoid arthritis.

doi: 10.1126/scisignal.adh1178

Figure Lengend Snippet: Fig. 8. Schematic model illustrating the proposed mechanism of CD4+ T cell pyroptosis in RA. In RA CD4+ T cells, ALOX5 is overexpressed, leading to an enhanced conversion of AA to LTB4, which subsequently increases ORAI3 abundance. ORAI3 oligomers form a Ca2+ channel that is activated by AA, and its increase results in increased Ca2+ influx into the cytoplasm. This surge in Ca2+ activates NLRP3, prompting it to form an inflammasome complex with the adapter protein ASC and pro- CASP1. This leads to autoprocessing of pro-CASP1 and the generation of active CASP1, which, in turn, cleaves GSDMD to yield GSDMD-N, inducing pyroptosis in CD4+ T cells.

Article Snippet: To explore the potential of ALOX5 pharmacologic inhibition in suppressing T cell pyroptosis, cells were treated with zileuton (MCE, HY- 14164) at 50 μM for 24 hours.

Techniques:

Journal: Communications Biology

Article Title: Targeting ALOX5 ameliorates renal tubular injury in ischemia–reperfusion-induced acute kidney injury via inhibition of ferroptosis

doi: 10.1038/s42003-025-08803-4

Figure Lengend Snippet: A The schematic graph showing the experimental process. B The heatmap showing the DEGs between the control and IRI-induced AKI groups in the chipset “ GSE192532 ,” which were also ferroptosis-related genes in the FerrDb database. C KEGG analysis showing the enrichment of the above DEGs in the ferroptosis pathway. D H&E and PAS staining showing the renal tubular injury in the IRI–AKI mouse model. E The IHC staining of Alox5 and the immunofluorescence (IF) staining to assess the up-regulated expression of Alox5 following IRI treatment. F Representative EM images of morphological changes in mitochondria cristae shape associated with ferroptosis (red arrow head) in renal tubular cells. Abbreviations: IHC immunohistochemistry, IF immunofluorescence, I/R ischemia-reperfusion, LM light microscopy, EM electron microscopy.

Article Snippet: Male Alox5 full knockout (Alox5 - / - ) mice (weighing 20-25 g, aged 8-10 weeks) were obtained from Cyagen Biology company (Suzhou, China).

Techniques: Control, Staining, Immunohistochemistry, Immunofluorescence, Expressing, Light Microscopy, Electron Microscopy

Journal: Communications Biology

Article Title: Targeting ALOX5 ameliorates renal tubular injury in ischemia–reperfusion-induced acute kidney injury via inhibition of ferroptosis

doi: 10.1038/s42003-025-08803-4

Figure Lengend Snippet: A The schematic graph showing the experimental process. B Increased serum creatinine and urea nitrogen levels in IRI mice, confirming renal function impairment following IRI treatment. n = 6 animals. C Arachidonic acid (AA) and its metabolic product 5-hydroxyeicosatetraenoic acid (5-HETE) detected by UHPLC-MS/MS. n = 6 animals. D The pathological examinations of kidney tissues, including the H&E and PAS staining showing the renal tubular injury, the IHC staining to detect the expression of ALOX5, the immunofluorescence staining to assess the existence of ALOX5, and the representative EM images of morphological changes in mitochondria cristae shape associated with ferroptosis (red arrow head) in renal tubular cells. E , F Western blot results showing the relative protein levels of GPX4 and xCT in the kidney tissues of mice. n = 3 independent experiments. Densitometric quantification normalized to β-actin is shown to the right of the immunoblots. All data are presented as mean ± standards errors. * p < 0.05. ** p < 0.01. Abbreviations: IHC immunohistochemistry, I/R ischemia–reperfusion, EM electron microscopy, WT wild-type.

Article Snippet: Male Alox5 full knockout (Alox5 - / - ) mice (weighing 20-25 g, aged 8-10 weeks) were obtained from Cyagen Biology company (Suzhou, China).

Techniques: Tandem Mass Spectroscopy, Staining, Immunohistochemistry, Expressing, Immunofluorescence, Western Blot, Electron Microscopy

Journal: Communications Biology

Article Title: Targeting ALOX5 ameliorates renal tubular injury in ischemia–reperfusion-induced acute kidney injury via inhibition of ferroptosis

doi: 10.1038/s42003-025-08803-4

Figure Lengend Snippet: A The schematic graph showing the experimental process. B Confocal microscopy results showing lipid peroxides detected by Liperfluo staining after ALOX5 knockdown or overexpression in HK2 receiving H/R treatment. C – F Western blot results showing the relative protein levels of GPX4 and xCT after ALOX5 knockdown in HK2 receiving H/R treatment. n = 3 independent experiments. G – J Western blot results showing the relative protein levels of GPX4 and xCT after ALOX5 overexpression in HK2 receiving H/R treatment. n = 3 independent experiments. Densitometric quantification normalized to β-actin is shown to the right of the immunoblots. All data are presented as mean ± standards errors. * p < 0.05. ** p < 0.01. Abbreviations: CTRL control, H/R hypoxia/reoxygenation, NC negative control, oe overexpression.

Article Snippet: Male Alox5 full knockout (Alox5 - / - ) mice (weighing 20-25 g, aged 8-10 weeks) were obtained from Cyagen Biology company (Suzhou, China).

Techniques: Confocal Microscopy, Staining, Knockdown, Over Expression, Western Blot, Control, Negative Control

Journal: Communications Biology

Article Title: Targeting ALOX5 ameliorates renal tubular injury in ischemia–reperfusion-induced acute kidney injury via inhibition of ferroptosis

doi: 10.1038/s42003-025-08803-4

Figure Lengend Snippet: A The schematic graph of the kinetic binding study using biolayer interferometry. B Molecular interaction experiment showing the combination between ALOX5 recombinant protein and BBR in different concentrations. C The steady-state analysis showing the degree of fitting of the molecular interaction curve (KD = 8.1E−05 ± 1.1E−05, R 2 = 0.9945). D Molecular docking results showing the combination between BBR and ALOX5. E The 2D image of the potential binding sites of ALOX5 to BBR. F The 3D image of the potential binding sites of ALOX5 to BBR. Abbreviations: BBR benzbromarone.

Article Snippet: Male Alox5 full knockout (Alox5 - / - ) mice (weighing 20-25 g, aged 8-10 weeks) were obtained from Cyagen Biology company (Suzhou, China).

Techniques: Binding Assay, Recombinant

Journal: Communications Biology

Article Title: Targeting ALOX5 ameliorates renal tubular injury in ischemia–reperfusion-induced acute kidney injury via inhibition of ferroptosis

doi: 10.1038/s42003-025-08803-4

Figure Lengend Snippet: A The schematic graph showing the experimental process. B , C Serum creatinine and blood urea nitrogen levels in the I/R mice model receiving BBR treatment. n = 6 animals. D The pathological examinations of kidney tissues, including the H&E and PAS staining showing the renal tubular injury and the IHC staining to detect the expression of ALOX5. E Representative EM images of morphological changes in mitochondria cristae shape associated with ferroptosis (red arrow head) in renal tubular cells. F – H Western blot results showing the relative protein levels of GPX4 and xCT after ALOX5 knockdown in the kidney tissue of I/R mice model receiving BBR treatment. n = 3 independent experiments. Densitometric quantification normalized to β-actin is shown to the right of the immunoblots. All data are presented as mean ± standards errors. * p < 0.05, ** p < 0.01. Abbreviations: BBR benzbromarone, DEX dexamethasone, EM electron microscopy, I/R ischemia–reperfusion.

Article Snippet: Male Alox5 full knockout (Alox5 - / - ) mice (weighing 20-25 g, aged 8-10 weeks) were obtained from Cyagen Biology company (Suzhou, China).

Techniques: Staining, Immunohistochemistry, Expressing, Western Blot, Knockdown, Electron Microscopy

Journal: Communications Biology

Article Title: Targeting ALOX5 ameliorates renal tubular injury in ischemia–reperfusion-induced acute kidney injury via inhibition of ferroptosis

doi: 10.1038/s42003-025-08803-4

Figure Lengend Snippet: A The schematic graph showing the experimental process. B CCK8 analysis results showing the cell viability of HK2 cells after BBR and H/R treatments. n = 5 independent experiments. C Cell viability of HK2 cells receiving treatment of RSL3 (1 μM), Fer1 (1 μM), and BBR (2 μM). n = 5 independent experiments. D Confocal microscopy results showing lipid peroxides detected by Liperfluo staining in HK2 cells receiving BBR (2 μM) and H/R treatments. E Confocal microscopy results showing lipid peroxides detected by Liperfluo staining in H/R-HK2 cells receiving BBR (2 μM) and ALOX5 interventions. F , G Western blot results showing the relative protein levels of ALOX5, GPX4, and xCT in HK2 cells receiving BBR (2 μM) and H/R treatments. n = 3 independent experiments. H – K Western blot results showing the relative protein levels of ALOX5, GPX4, and xCT in H/R-HK2 cells receiving BBR (2 μM) and ALOX5 interventions. n = 3 independent experiments. Densitometric quantification normalized to β-actin is shown to the right of the immunoblots. All data are presented as mean ± standards errors. * p < 0.05; ** p < 0.01. Abbreviations: BBR benzbromarone, DEX dexamethasone, H/R hypoxia/reoxygenation.

Article Snippet: Male Alox5 full knockout (Alox5 - / - ) mice (weighing 20-25 g, aged 8-10 weeks) were obtained from Cyagen Biology company (Suzhou, China).

Techniques: Confocal Microscopy, Staining, Western Blot

Journal: Communications Biology

Article Title: Targeting ALOX5 ameliorates renal tubular injury in ischemia–reperfusion-induced acute kidney injury via inhibition of ferroptosis

doi: 10.1038/s42003-025-08803-4

Figure Lengend Snippet: A The schematic graph showing the experimental process. B The pathological examinations of adjacent non-tumorous kidney tissue in one patient with renal tumor and kidney tissue obtained via renal biopsy in one patient with acute tubular necrosis, respectively, including the H&E staining showing the renal tubular injury, the IHC and IF staining to detect the expression of ALOX5, and the representative EM images of morphological changes in mitochondria cristae shape associated with ferroptosis (red arrow head) in renal tubular cells. Abbreviations: ATN acute tubular necrosis, IF immunofluorescence, IHC immunohistochemistry, EM electron microscopy, LM light microscopy.

Article Snippet: Male Alox5 full knockout (Alox5 - / - ) mice (weighing 20-25 g, aged 8-10 weeks) were obtained from Cyagen Biology company (Suzhou, China).

Techniques: Staining, Expressing, Immunofluorescence, Immunohistochemistry, Electron Microscopy, Light Microscopy

Journal: Oxidative Medicine and Cellular Longevity

Article Title: 5-Lipoxygenase Inhibition Protects Retinal Pigment Epithelium from Sodium Iodate-Induced Ferroptosis and Prevents Retinal Degeneration

doi: 10.1155/2022/1792894

Figure Lengend Snippet: Primers of mouse genes for real-time PCR.

Article Snippet: To knockdown ALOX5 , ARPE-19 cells were transfected for 72 h in 24-well plates with three unique 27mer siRNA duplexes specific for human ALOX5 (OriGene, SR319325, 10 nM), or with scrambled siRNA (OriGene SR30004, 10 nM) as negative control, using the Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher) according to the manufacturer's protocol.

Techniques:

Journal: Oxidative Medicine and Cellular Longevity

Article Title: 5-Lipoxygenase Inhibition Protects Retinal Pigment Epithelium from Sodium Iodate-Induced Ferroptosis and Prevents Retinal Degeneration

doi: 10.1155/2022/1792894

Figure Lengend Snippet: Inhibition of lipoxygenase-5 (5-LOX) protected ARPE-19 cells from NaIO 3 -induced cell death by ferroptosis. (a) Arachidonic acid (ARA, black arrow) and 5-hydroxyeicosatetraenoic acid (5-HETE, white arrow) were detected with HPLC-MS/MS analysis. CCK-8 was used to analyze cell viability following treatments (b–f). Cell viability of ARPE-19 following treatment with (b) nonlethal concentration of NaIO 3 (10 mM, 20 h) and different concentrations of arachidonic acid, (c) toxic NaIO 3 (35 mM, 20 h) and different concentrations of 5-LOX inhibitor Zileuton ( n = 6 per treatment condition), and (d) other types of LOX inhibitors (75 μ M Zileuton, 15 μ M Mk-886, 3 μ M ML355, 500 nM BLX-3887, n = 6 per treatment condition). (e) The effect of NaIO 3 (35 mM, 20 h) with ALOX5 knockdown-ARPE-19 cell. (f) Effect of combined treatment with suboptimal concentrations of Zileuton (40 μ M) and DFO (25 μ M) on NaIO 3 (35 mM, 20 h)-induced ferroptosis. Abbreviations: HPLC-MS/MS: high-performance liquid chromatography with tandem mass spectrometry; PBS: phosphate-buffered saline; sc: scramble; KD: knockdown.

Article Snippet: To knockdown ALOX5 , ARPE-19 cells were transfected for 72 h in 24-well plates with three unique 27mer siRNA duplexes specific for human ALOX5 (OriGene, SR319325, 10 nM), or with scrambled siRNA (OriGene SR30004, 10 nM) as negative control, using the Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher) according to the manufacturer's protocol.

Techniques: Inhibition, Tandem Mass Spectroscopy, CCK-8 Assay, Concentration Assay, Knockdown, High Performance Liquid Chromatography, Mass Spectrometry, Saline

Journal: Oxidative Medicine and Cellular Longevity

Article Title: 5-Lipoxygenase Inhibition Protects Retinal Pigment Epithelium from Sodium Iodate-Induced Ferroptosis and Prevents Retinal Degeneration

doi: 10.1155/2022/1792894

Figure Lengend Snippet: Inhibition of lipoxygenase-5 (5-LOX) protected ARPE-19 cells from cell death induced by conventional ferroptosis inducers. Cell viability of ARPE-19 cells following ferroptosis induction with (a, b) Erastin (24 h, n = 6 per treatment condition), (c) combined treatment with ferrous ion (FeSO 4 100 μ M, 8 h) and tBHP (250 μ M, 8 h, n = 6 per treatment condition), and (d) RSL3 (1 μ M, 20 h). Abbreviations: tBHP: tert-Butyl hydroperoxide.

Article Snippet: To knockdown ALOX5 , ARPE-19 cells were transfected for 72 h in 24-well plates with three unique 27mer siRNA duplexes specific for human ALOX5 (OriGene, SR319325, 10 nM), or with scrambled siRNA (OriGene SR30004, 10 nM) as negative control, using the Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher) according to the manufacturer's protocol.

Techniques: Inhibition

Journal: Oxidative Medicine and Cellular Longevity

Article Title: 5-Lipoxygenase Inhibition Protects Retinal Pigment Epithelium from Sodium Iodate-Induced Ferroptosis and Prevents Retinal Degeneration

doi: 10.1155/2022/1792894

Figure Lengend Snippet: Inhibition of lipoxygenase-5 (5-LOX) reduced NaIO 3 -induced oxidative stress in ARPE-19 cells. (a, b) BODIPY C11 staining lipid peroxidation ( n = 6, 35 mM NaIO 3 ,15 h, Zileuton 75 μ M). (c) FerroOrange staining for intracellular ferrous ions (75 μ M Zileuton, n = 12). (d) Ferroptosis-associated protein xCT detected with western blot analysis (18 mM NaIO 3 for 15 h, n = 3). (e) Oxidative stress-induced DNA damage demonstrated with 8-OHdG staining ( n = 4, 35 mM NaIO 3 , 75 μ M Zileuton, 15 h). Scale bar = 50 μ m. Abbreviations: xCT: cystine-glutamate antiporter; 8-OHdG: 8-hydroxy-2′-deoxyguanosine; Zil: Zileuton.

Article Snippet: To knockdown ALOX5 , ARPE-19 cells were transfected for 72 h in 24-well plates with three unique 27mer siRNA duplexes specific for human ALOX5 (OriGene, SR319325, 10 nM), or with scrambled siRNA (OriGene SR30004, 10 nM) as negative control, using the Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher) according to the manufacturer's protocol.

Techniques: Inhibition, Staining, Western Blot

Journal: Discover Oncology

Article Title: Lipid metabolism-related genes correlate with immune microenvironment and regulate the efficacy of immunotherapy via ferroptosis in melanoma

doi: 10.1007/s12672-025-04163-x

Figure Lengend Snippet: Screening and functional validation of differentially-expressed ferroptosis-related genes. (A) Venn diagrams of the differentially-expressed ferroptosis-related genes that have prognostic potential. (B) The expression of ALOX5, ACSL4, ACACA, ABCC1 in the two subgroups in training cohort, as well as Kaplan-Meier analysis of the survival of patients in the two subgroups. (C) The knockdown efficiency of ASCL4 and ALOX5 in melanoma cells, as well as cell viability of melanoma cells in response to RSL3 treatment. (D) Lipid ROS levels of indicated cells measured by flow cytometry using C11-BODIPY. Data represent the mean ± SEM of triplicates. P value was calculated by two-tailed Student’s t-test and log-rank tests. ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns, non-significant. n (cluster 1) = 208, n (cluster 2) = 246. n (low) = 227, n (high) = 227

Article Snippet: The primary antibodies and dilutions for western blotting (WB) and immunofluorescence (IF) staining analysis are listed below: ACSL4 (22401-1-AP, Proteintech, Wuhan, China; 1:100 for IHC, 1:100 for IF and 1:5 000 for WB), ALOX5 (10021-1-Ig, Proteintech; 1:400 for IHC, and 1:500 for WB), PTGS2 (66351-1-Ig, Proteintech; 1:400 for IF), CD8α(ab22378,, Abcam; 1:300 for IF), ACTIN (66009-1-Ig, Proteintech; 1:10 000 for WB).

Techniques: Functional Assay, Biomarker Discovery, Expressing, Knockdown, Flow Cytometry, Two Tailed Test

Journal: Journal of Translational Medicine

Article Title: ERO1A promotes the proliferation, migration and invasion of bladder cancer through ALOX5 mediated activation of JAK-STAT signaling pathway

doi: 10.1186/s12967-025-07613-w

Figure Lengend Snippet: ERO1A can inhibit the phosphorylation level of ALOX5. ( A ) The binding sites between ERO1A and ALOX5 were predicted using molecular docking technology. ( B ) The interaction between ERO1A and ALOX5 in bladder cancer cells was verified by co-immunoprecipitation. ( C ) Co-IP method was used to detect the binding of ERO1A and ALOX5 in 293T cells. ( D ) The mutant R154A and C208A were transfected into bladder cancer cells; co-immunoprecipitation experiments were conducted to detect the functional fragments. R154A: The arginine at position 154 was replaced by alanine; C208A: The cysteine at position 208 was replaced by alanine. ( E ) The co-localization of ERO1A and ALOX5 in bladder cancer cells was observed by immunofluorescence, scale bar: 10 μm. ( F-H ) The effect of silencing ERO1A expression on ALOX5 expression. ( I-K ) The effect of ERO1A overexpression on ALOX5 expression. Student’s t-test (two groups) and one-way analysis of variance (more than two groups) were used to analyze the differences between groups. All assays were independently repeated three times. All data are expressed as mean ± standard deviation (SD). ** P < 0.01, *** P < 0.001

Article Snippet: p-ALOX5 , ABclonal , AP0573 , Rabbit , WB , 1:1000,1:50.

Techniques: Phospho-proteomics, Binding Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Mutagenesis, Transfection, Functional Assay, Immunofluorescence, Expressing, Over Expression, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: ERO1A promotes the proliferation, migration and invasion of bladder cancer through ALOX5 mediated activation of JAK-STAT signaling pathway

doi: 10.1186/s12967-025-07613-w

Figure Lengend Snippet: ALOX5 can counteract the effects of ERO1A on the JAK-STAT signaling pathway and the autophagy signaling pathway. ( A-C ) The western blotting was used to detect the effects of ALOX5 overexpression on the expression of proteins related to the JAK-STAT signaling pathway and the autophagy signaling pathway. ( D-F ) The western blotting was used to detect the effects of ALOX5 gene silencing on the expression of proteins related to the JAK-STAT signaling pathway and the autophagy signaling pathway. ( G-I ) The western blotting showed that silencing ALOX5 could partially counteract the inhibitory effect of ERO1A knockdown on the JAK-STAT signaling pathway and the promoting effect on the autophagy signaling pathway. ( J-L ) The western blotting indicated that the overexpression of ALOX5 could partially counteract the activation of the JAK-STAT signaling pathway and the inhibition of the autophagy signaling pathway induced by ERO1A overexpression. Student’s t-test (two groups) and one-way analysis of variance (more than two groups) were used to analyze the differences between groups. All assays were independently repeated three times. All data are expressed as mean ± standard deviation (SD). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: p-ALOX5 , ABclonal , AP0573 , Rabbit , WB , 1:1000,1:50.

Techniques: Western Blot, Over Expression, Expressing, Knockdown, Activation Assay, Inhibition, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: ERO1A promotes the proliferation, migration and invasion of bladder cancer through ALOX5 mediated activation of JAK-STAT signaling pathway

doi: 10.1186/s12967-025-07613-w

Figure Lengend Snippet: ERO1A regulates the JAK/STAT/autophagy signaling axis by modulating the phosphorylation of ALOX5, thereby promoting the malignant development of bladder cancer. ( A ) The effect of ERO1A gene silencing on tumor volume. Each group n = 6 (6 nude mice). ( B ) Subcutaneous tumor masses of mice in the shNC group and shERO1A group. Each group n = 6 (6 nude mice). ( C ) The effect of ERO1A gene silencing on tumor weight. Each group n = 6 (6 nude mice). ( D-E ) Detection of the expression levels of JAK-STAT-related proteins and autophagy-related proteins by western blotting. ( F ) Detection of the expression of related indicators in the tumors of the two groups by immunohistochemistry, scale: 200 μm. ( G ) HE staining demonstrated the number of lung metastases in tumor samples from the shNC and shERO1A groups, scale: 200 μm. Each group n = 6 (6 nude mice). Student’s t-test (two groups) and one-way analysis of variance (more than two groups) were used to analyze the differences between groups. All assays were independently repeated three times. All data are expressed as mean ± standard deviation (SD). * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: p-ALOX5 , ABclonal , AP0573 , Rabbit , WB , 1:1000,1:50.

Techniques: Phospho-proteomics, Expressing, Western Blot, Immunohistochemistry, Staining, Standard Deviation

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 1 Alox5-5-HETE axis is upregulated in gastric cancer tissues. (A) Scatter plot of Alox5 protein level in paired normal and tumor tissues obtained from 52 patients with gastric tumor. (B) Representative immunohistochemistry analysis of normal and tumor gastric tissues from patient#23 performed by Alox5 staining. (C) Scatter plot of 5-HETE level in paired normal and tumor tissues obtained from 52 patients with gastric tumor. Alox5 and 5-HETE were assessed using tissue homogenates and quantified using ELISA assay. (D) Relative tumor/normal ratio value of Alox5 and 5-HETE levels in individual gastric cancer patients. Alox5 or 5-HETE level in normal tissues were set as 1 (indicated by a red line)

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Immunohistochemistry, Staining, Enzyme-linked Immunosorbent Assay

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 2 ALOX5 overexpression promotes cell growth and protects gastric cancer cells from chemotherapeutic agents-induced toxicity. (A and B) Analysis of proliferation of AGS and N87 cells after ALOX5 overexpression performed by BrdU labeling. (C and D) Analysis of colony formation of AGS and N87 cells after ALOX5 overexpression. (E and F) Analysis of migration of AGS and N87 cells after ALOX5 overexpression. Analysis of proliferation (G and H) and apoptosis (I and J) after treatment of 5-FU and cisplatin in ALOX5-overexpressing AGS and N87 cells. Results are presented as relative to control. Proliferation and apoptosis assays were assessed after 72 h of drug treatment. 5-FU at 200 nM and cisplatin at 300 nM were used. *p < 0.05, compared to p-Vector. #p < 0.05, compared to cisplatin or 5-FU alone

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Over Expression, Labeling, Migration, Control, Plasmid Preparation

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 4 ALOX5 knockdown inhibits gastric cancer and enhances the toxicity of chemotherapeutic agents. Analysis of proliferation (A and B) and apoptosis (C and D) in AGS and N87 cells after ALOX5 knockdown, and in the presence of 5-FU and cisplatin. ALOX5 knockdown significantly enhances the anti-proliferative and pro-apoptotic effects of 5-FU and cisplatin in gastric cancer cells. 5-FU at 50 nM and cisplatin at 50 nM were used. Proliferation and apoptosis assays were assessed after 72 h of drug treatment. Results are presented as relative to control. *p < 0.05, compared to control. #p < 0.05, compared to cisplatin or 5-FU

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Knockdown, Control

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 6 ALOX5 knockdown inhibits ERK in gastric cancer cells. Representative western blot photo (A) and quantification analysis by Image J (B) of p-Erk, p-p90RSK, p-Akt, pol II S5, Mcl-1, Bim, and Bcl levels in N87 cells after ALOX5 knockdown. Scr siRNA value was set as 1 and indicated as line. *p < 0.05, compared to Scr siRNA

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Knockdown, Western Blot

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 5 Alox5 inhibitors suppress gastric cancer and enhance the toxicity of chemotherapeutic agents. (A and B) Proliferation level of AGS and N87 cells after zileuton and AA861 treatment in the presence of 5-FU and cisplatin, as evaluated by BrdU labeling. (C and D) Proliferation level of AGS and N87 cells after zileuton and AA861 treatment in the presence of 5-FU and cisplatin, as evaluated by TUNEL assay. Zileuton and AA861 significantly enhance the anti-proliferative and pro-apoptotic effects of 5-FU and cisplatin in gastric cancer cells. 5-FU at 50 nM and cisplatin at 50 nM were used. Zileuton at 100 μM and 200 μM, AA86 at 30 μM and 60 μM were used in proliferation and apoptosis assays, respectively. Proliferation and apoptosis assays were assessed after 72 h of drug treatment. *p < 0.05, compared to control. #p < 0.05, compared to cisplatin or 5-FU

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Labeling, TUNEL Assay, Control

Journal: Cancer medicine

Article Title: ALOX5-5-HETE promotes gastric cancer growth and alleviates chemotherapy toxicity via MEK/ERK activation.

doi: 10.1002/cam4.4066

Figure Lengend Snippet: FIGURE 7 ALOX5-5-HETE axis activates ERK in gastric cancer cells. (A) Representative western blot photo of p-Erk, p-p90RSK, p- Akt, Mcl-1, Bim, and Bcl-2 in N87 cells after ALOX5 overexpression or 5-HETE addition. (B) Proliferation level of treatment of U0126 and LY2780301 in ALOX5-overexpressing N87 cells. (C) Proliferation level of treatment of U0126 and LY2780301 in N87 cells in the presence of 5-HETE. U0126 (MEK inhibitor, 10 μM) but not LY2780301 (Akt inhibitor, 10 μM) significantly reveres the pro-proliferative effect induced by ALOX5 overexpression and addition of 5-HETE. Inhibitors were added to the cells at 48h post-transfection. *p < 0.05, compared to control; ns, not significant

Article Snippet: ALOX5- overexpressing plasmid (pCMV6ALOX5, RG217259) and vector control plasmid (pCMV6AC, PS100010) were obtained from OriGene.

Techniques: Western Blot, Over Expression, Transfection, Control

Journal: Clinical and Translational Medicine

Article Title: Gut microbiota of miR‐30a‐5p‐deleted mice aggravate high‐fat diet‐induced hepatic steatosis by regulating arachidonic acid metabolic pathway

doi: 10.1002/ctm2.70035

Figure Lengend Snippet: The expression level of proteins involved in the arachidonic acid metabolic pathway in liver tissues. Immunohistochemistry staining images of the expression levels of ALOX5 (A), ALOX12 (B), ALOX15 (C), COX‐1 (D), and COX‐2 (E) in liver tissues of WT or KO mice treatment with ND or HFD (scale bar: 100 µm). (F) The quantitative results of immunohistochemistry are determined by statistical average optical density (AOD). (G) The mRNA levels of ALOX5, ALOX12, ALOX15, COX‐1, and COX‐2 in liver tissue of WT or KO mice treated with ND or HFD were detected by real‐time PCR. Data are expressed as mean ± SD and one‐way ANOVA with Tukey's test was used for multiple comparisons, NS showed no significant difference, * p < .05, ** p < .01, *** p < .001.

Article Snippet: Sections were incubated with primary antibodies against ALOX5 (Cat#: A2158, Abclonal, Wuhan, China), ALOX12 (Cat#: A14703, Abclonal, Wuhan, China), ALOX15 (Cat#: A6864, Abclonal, Wuhan, China), COX1 (Cat#: A7341, Abclonal, Wuhan, China), COX2 (Cat#: A3560, Abclonal, Wuhan, China).

Techniques: Expressing, Immunohistochemistry, Staining, Real-time Polymerase Chain Reaction

Journal: Clinical and Translational Medicine

Article Title: Gut microbiota of miR‐30a‐5p‐deleted mice aggravate high‐fat diet‐induced hepatic steatosis by regulating arachidonic acid metabolic pathway

doi: 10.1002/ctm2.70035

Figure Lengend Snippet: AAV‐OE reversed the HFD‐KO mice induced hepatic steatosis and COX/LOX pathways. (A) We injected 100 µL/each AAV‐NC or AAV‐OE into miR‐30a‐5p −/− mice through the tail vein. Two weeks after injection, the mice began HFD for 8 weeks. (B) Representative images of H&E staining and Oil Red O of liver tissues in KO + HFD mice treatment with AAV‐NC or AAV‐OE ( n = 5, scale bar: 100 µm). (C) The levels of total cholesterol, triglyceride, HDL‐c, and LDL‐c in the serum of KO + HFD mice treatment with AAV‐NC or AAV‐OE ( n = 5). (D) The levels of ALT, AST, total bile acid, and cholinesterase in the serum of KO + HFD mice treatment with AAV‐NC or AAV‐OE ( n = 5). (E) ALOX5 (F), ALOX12 and (G) COX‐2 in liver tissues of KO + HFD mice treatment with AAV‐NC or AAV‐OE ( n = 5, scale bar: 100 µm). (H‐I) The mRNA levels of ALOX5, ALOX12, and COX‐2 in liver tissue of KO + HFD mice treatment with AAV‐NC or AAV‐OE ( n = 5). Data are expressed as mean ± SD and one‐way ANOVA with Tukey's test was used for multiple comparisons, NS showed no significant difference, * p < .05, ** p < .01.

Article Snippet: Sections were incubated with primary antibodies against ALOX5 (Cat#: A2158, Abclonal, Wuhan, China), ALOX12 (Cat#: A14703, Abclonal, Wuhan, China), ALOX15 (Cat#: A6864, Abclonal, Wuhan, China), COX1 (Cat#: A7341, Abclonal, Wuhan, China), COX2 (Cat#: A3560, Abclonal, Wuhan, China).

Techniques: Injection, Staining