Journal: Experimental & Molecular Medicine

Article Title: Regulation of PKM2 expression and function by GLIS3 during metabolic reprogramming in polycystic kidneys

doi: 10.1038/s12276-026-01676-5

Figure Lengend Snippet: a , Representative images comparing whole kidney size between Glis3-KO2 kidneys treated with vehicle or compound 3K are shown. b , Violin plot depicting the KW/BW ratio (%) for WT and Glis3 -KO2 mice treated with vehicle or compound 3K. n ≥ 10; **** P < 0.0001. c , Representative hematoxylin and eosin-scanned images of kidney sections from Glis3 -KO2 kidneys treated with vehicle or compound 3K. Bars indicate 1 mm. d , Comparison of cystic index between Glis3 -KO2 kidneys treated with vehicle or compound 3K. Cystic index represents the percentage of renal tissue occupied by cysts. Data are presented as mean ± s.e.m., n = 6 . ** P < 0.01. e , Comparison of renal cyst size (mm 2 ) between kidneys from Glis3 -KO2 mice treated with vehicle or compound 3K. Data are presented as mean ± s.e.m., n = 6; * P < 0.05. f , Comparison of the number of cysts per kidney section between Glis3 -KO2 kidneys treated with vehicle or compound 3K. Data are presented as mean ± s.e.m., n = 8; ** P < 0.01. g , Comparison of serum creatinine levels (mg/dl) between WT and Glis3 -KO2 mice treated with vehicle or compound 3K. h , RT–qPCR analysis of Pkm2 , c-Myc , Hk2 , Havcr1 and Lcn2 between WT and Glis3 -KO2 mice treated with vehicle or compound 3K. Data are presented as mean ± s.e.m., n ≥ 5; **** P < 0.0001; *** P < 0.001; ** P < 0.01; * P < 0.05. i , Schematic illustration depicting the relationship between loss of GLIS3 function, regulation of PKM2 and cystogenesis. GLIS3 deficiency enhances Pkm gene expression in kidneys with a preferential increase in the Pkm2 isoform. Increased PKM2 phosphorylation at S37 and Y105 promotes dimer formation. PKM2-S37 phosphorylation is facilitated by increased pERK1/2 levels. Together, these events promote glycolysis, cell proliferation and cystogenesis in GLIS3-deficient kidneys. Figure 6i was created using BioRender.com.

Article Snippet: To examine the effect of PKM2 inhibition on cyst formation, PND7 Glis3 -Pax8Cre mice were treated intraperitoneally with the PKM2 inhibitor, compound 3K (S8616; Selleckchem Chemicals) (10 mg/kg) or vehicle control (10% dimethylsulfoxide (DMSO), 90% corn oil) every 24 h for 7 days.

Techniques: Comparison, Quantitative RT-PCR, Gene Expression, Phospho-proteomics

Journal: Experimental & Molecular Medicine

Article Title: Regulation of PKM2 expression and function by GLIS3 during metabolic reprogramming in polycystic kidneys

doi: 10.1038/s12276-026-01676-5

Figure Lengend Snippet: a , Immunoblot analysis of PKM2 protein levels in WT and Glis3 -KO2 RECS 3 days after siRNA-mediated PKM2-KD. Protein expression was quantified by densitometric analysis. Data are presented as mean ± s.e.m., n = 5; *** P < 0.001; ** P < 0.01. b , Analysis of lactate production in media from primary WT and Glis3 -KO2 RECs with or without PKM2-KD. c , Glycolytic rate was measured in primary WT and Glis3 -KO2 REC mice with or without PKM2-KD using a Seahorse analyzer after sequential injections of rotenone/antimycin A and 2-DG. d , e , Basal ( d ) and compensatory ( e ) glycolysis were calculated and plotted ( n = 3). f , Representative images of WT and Glis3 -KO2 REC spheroids with and without PKM2-KD at 5 and 9 days. Bars indicate 50 μm. g , Violin plot showing the size (µm) distribution of the spheroids generated at day 5 from WT and Glis3 -KO2 RECs with or without PKM2-KD ( n ≥ 4). Each data point represents an individual spheroid measurement. * P < 0.05, ** P < 0.01, *** P < 0.001. h , Spheroid images at day 5 were taken using the EVOS M7000, and spheroid diameter and number were analyzed using ImageJ and plotted according to size distribution—either 30–50, 50–100 or >100 μm. Total indicates the number of spheroids analyzed in each group ( n = 39–164). i , Violin plot showing the size (µm) distribution of the spheroids generated at day 9 from WT and Glis3 -KO2 REC mice with or without PKM2-KD ( n ≥ 4). Each data point represents an individual spheroid measurement. * P < 0.05, ** P < 0.01, *** P < 0.001. j , Day 10 spheroid size distribution—either 30–50, 50–100 or >100 μm. Total indicates the number of spheroids analyzed in each group ( n = 60–191). k , Representative image of the size of WT and Glis3 -KO2 REC spheroids 5 days following treatment with vehicle (0.1% DMSO) or compound 3K (1 μM). Bars indicate 50 μm. l , Violin plot showing the size (µm) distribution of the spheroids generated from the RECs of WT and Glis3 -KO2 kidneys ( n ≥ 4). Each data point represents an individual spheroid measurement. **** P < 0.0001. m , Size distribution—30–50, 50–100 or >100 μm of WT and Glis3 -KO2 REC spheroids with or without PKM2 inhibition. Total indicates the number of spheroids analyzed in each group (n = 39–164).

Article Snippet: To examine the effect of PKM2 inhibition on cyst formation, PND7 Glis3 -Pax8Cre mice were treated intraperitoneally with the PKM2 inhibitor, compound 3K (S8616; Selleckchem Chemicals) (10 mg/kg) or vehicle control (10% dimethylsulfoxide (DMSO), 90% corn oil) every 24 h for 7 days.

Techniques: Western Blot, Expressing, Generated, Inhibition

Journal: International Journal of Biological Sciences

Article Title: ARID1A deficiency activates OSM-STAT3 axis in endometrial cancer, creating vulnerability to JAK/STAT3 inhibition

doi: 10.7150/ijbs.129142

Figure Lengend Snippet: Identification of synthetic lethality of ARID1A and JAK/STAT3 in endometrial cancer through drug library screening. A. Schematic illustration of the synthetic lethality screenings with the Epigenetics Drug Library and the Kinase Inhibitor Library. B. Immunoblot analysis showing ARID1A knockout in A1 and B7 single clone. C. Drugs with selectivity index (SI)≥2 were selected as synthetic lethality candidates. SI=IC50 ARID1A+/+ /IC50 ARID1A-/- . D. Synthetic lethality in HEC1B-ARID1A -/- cell treated with stattic (D, F) and gandotinib (E, G) for 72h. The cell images were taken with IncuCyte ZOOM. Scale bar, 300 μm. H. Immunoblot analysis validated the ARID1A overexpression in HEC1B-ARID1A -/- cell. I. Overexpressed ARID1A rescued the synthetic lethality mediated by stattic. Data are mean±sd. * P <0.05, ** P <0.01. J-K. HEC1B ARID1A isogenic cell pair were transfected with STAT3-siRNA for 72 h. J. The quantitative analysis of synthetic lethality induced by STAT3-siRNA. Data are mean±sd. *** P <0.001. K. The growth curve of HEC1B ARID1A isogenic cell pair treated with STAT3-siRNA. **** P <0.0001. Two-way ANOVA test. L- N. IC50 test treated with stattic and gandotinib in endometrial cancer cell line. Data are mean±sd. * P <0.05.

Article Snippet: Epigenetics Drug Library and the Kinase Inhibitor Library screening were purchased from Selleck Chemicals.

Techniques: Drug discovery, Western Blot, Knock-Out, Over Expression, Transfection

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: We used the Octet RED96e system (ForteBio, USA) to identify small molecules that interact with ALOX5 in commercial compound libraries (Selleck L3300 Compound Libraries) by compound screening.

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: We used the Octet RED96e system (ForteBio, USA) to identify small molecules that interact with ALOX5 in commercial compound libraries (Selleck L3300 Compound Libraries) by compound screening.

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: We used the Octet RED96e system (ForteBio, USA) to identify small molecules that interact with ALOX5 in commercial compound libraries (Selleck L3300 Compound Libraries) by compound screening.

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: We used the Octet RED96e system (ForteBio, USA) to identify small molecules that interact with ALOX5 in commercial compound libraries (Selleck L3300 Compound Libraries) by compound screening.

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: We used the Octet RED96e system (ForteBio, USA) to identify small molecules that interact with ALOX5 in commercial compound libraries (Selleck L3300 Compound Libraries) by compound screening.

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: We used the Octet RED96e system (ForteBio, USA) to identify small molecules that interact with ALOX5 in commercial compound libraries (Selleck L3300 Compound Libraries) by compound screening.

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: We used the Octet RED96e system (ForteBio, USA) to identify small molecules that interact with ALOX5 in commercial compound libraries (Selleck L3300 Compound Libraries) by compound screening.

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