Journal: iScience

Article Title: GUTK induces apoptosis in reactivating quiescent prostate cancer cells via Aurora A-mediated stabilization of SOD2

doi: 10.1016/j.isci.2026.115739

Figure Lengend Snippet: SOD2 knockdown rescues GUTK-induced apoptosis and ΔΨm loss in reactivating quiescent PCa cells (A and B) Immunoblot analysis of SOD2 protein levels in quiescent LNCaP (A) and DU145 (B) cells treated with 20 μM GUTK for 12–48 h during cell cycle re-entry. β-actin and GAPDH served as loading controls. Quantification of relative protein levels is shown in the right panel. (C and D) Validation of SOD2 knockdown by immunoblot analysis in LNCaP (C) and DU145 (D) cells transfected with shSOD2. β-actin or GAPDH served as loading controls. Quantification of protein levels is shown in the right panel. (E and F) LNCaP shSOD2 and DU145 shSOD2 cells were treated with or without DOX to induce SOD2 knockdown. Apoptosis was assessed by Annexin V-APC/PI staining and flow cytometry in LNCaP shSOD2 (E) and DU145 shSOD2 (F) cells treated with DMSO or 20 μM GUTK for the indicated times during cell cycle re-entry. Quantification of apoptotic cell percentage is shown in the right panel. (G and H) ΔΨm assessment by JC-1 staining and flow cytometry in quiescent LNCaP shSOD2 (G) and DU145 shSOD2 (H) cells treated with either DMSO or 20 μM GUTK for the indicated times during cell cycle re-entry. Quantification of cells with low ΔΨm is shown in the right panel. Cont: proliferating control cells; Qsct: quiescent cells. ΔΨm: mitochondrial membrane potential. Data are presented as mean ± SD from three independent experiments. ∗ p < 0.05 and ∗∗∗ p < 0.001 versus indicated group.

Article Snippet: Sections were subjected to hematoxylin and eosin (H&E) staining or immunohistochemical staining using primary antibodies against Ki-67 (ab16667, Abcam, Shanghai, China), SOD2 (ET1701-54, HUABIO, Hangzhou, China), Cleaved Caspase 3 (#9661, Cell Signaling Technology, Danvers, USA) and Aurora A (ET1609-22, HUABIO, Hangzhou, China).

Techniques: Knockdown, Western Blot, Biomarker Discovery, Transfection, Staining, Flow Cytometry, Control, Membrane

Journal: iScience

Article Title: GUTK induces apoptosis in reactivating quiescent prostate cancer cells via Aurora A-mediated stabilization of SOD2

doi: 10.1016/j.isci.2026.115739

Figure Lengend Snippet: SOD2 overexpression exacerbates GUTK-induced apoptosis and ΔΨm loss in reactivating quiescent PCa cells (A and B) Immunoblot validation of SOD2 overexpression in LNCaP (A) and DU145 (B) cells stably transfected with empty vector (EV) or SOD2 expression construct (SOD2 OE). GAPDH served as the loading control. Quantification of relative protein levels is shown in the right panel. (C and D) Annexin V-FITC/PI staining and flow cytometry analysis of EV control or SOD2 OE LNCaP (C) and DU145 (D) cells treated with or without 20 μM GUTK for the indicated times during cell cycle re-entry. Quantification of apoptotic cell percentages is shown in the right panel. (E and F) ΔΨm analysis by JC-1 staining and flow cytometry in EV control or SOD2 OE LNCaP (E) and DU145 (F) cells treated with or without 20 μM GUTK for the indicated times during cell cycle re-entry. Quantification of cells with low ΔΨm is shown in the right panel. ΔΨm: mitochondrial membrane potential. Data are presented as mean ± SD from three independent experiments. ∗∗∗ p < 0.001 versus indicated group.

Article Snippet: Sections were subjected to hematoxylin and eosin (H&E) staining or immunohistochemical staining using primary antibodies against Ki-67 (ab16667, Abcam, Shanghai, China), SOD2 (ET1701-54, HUABIO, Hangzhou, China), Cleaved Caspase 3 (#9661, Cell Signaling Technology, Danvers, USA) and Aurora A (ET1609-22, HUABIO, Hangzhou, China).

Techniques: Over Expression, Western Blot, Biomarker Discovery, Stable Transfection, Transfection, Plasmid Preparation, Expressing, Construct, Control, Staining, Flow Cytometry, Membrane

Journal: iScience

Article Title: GUTK induces apoptosis in reactivating quiescent prostate cancer cells via Aurora A-mediated stabilization of SOD2

doi: 10.1016/j.isci.2026.115739

Figure Lengend Snippet: GUTK downregulates Aurora A expression during the reactivation of quiescent PCa cells (A and B) RT-PCR analysis of SOD2 mRNA levels in quiescent LNCaP (A) and DU145 (B) cells treated with or without 20 μM GUTK for the indicated times during cell cycle re-entry. (C and D) Quiescent LNCaP and DU145 cells were induced to re-enter the cell cycle and simultaneously exposed to 10 μM MG132 or DMSO for the indicated times. Immunoblot analysis of SOD2 protein was performed in quiescent LNCaP (C) and DU145 (D) cells. GAPDH served as the loading control. Quantification of relative protein levels is shown in the lower panel. (E and F) Quiescent LNCaP and DU145 cells were induced to re-enter the cell cycle for 6 h and then exposed to 50 μM CHX or DMSO for the indicated times. Immunoblot analysis of SOD2 and Aurora A proteins was performed in quiescent LNCaP (E) and DU145 (F) cells. β-actin or GAPDH served as loading control. Quantification of relative protein levels is shown in the lower panel. (G and H) Immunoblot analysis of Aurora A protein levels in quiescent LNCaP (G) and DU145 (H) cells treated with 20 μM GUTK for the indicated times during cell cycle re-entry. β-actin served as loading control. Quantification of relative protein levels is shown in the lower panel. (I and J) RT-PCR analysis of Aurora A mRNA levels in quiescent LNCaP (I) and DU145 (J) cells treated with 20 μM GUTK for the indicated times during cell cycle re-entry. Cont: proliferating control cells; Qsct: quiescent cells; CHX: cycloheximide. Data are presented as mean ± SD from three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus indicated group. ns: not significant.

Article Snippet: Sections were subjected to hematoxylin and eosin (H&E) staining or immunohistochemical staining using primary antibodies against Ki-67 (ab16667, Abcam, Shanghai, China), SOD2 (ET1701-54, HUABIO, Hangzhou, China), Cleaved Caspase 3 (#9661, Cell Signaling Technology, Danvers, USA) and Aurora A (ET1609-22, HUABIO, Hangzhou, China).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: iScience

Article Title: GUTK induces apoptosis in reactivating quiescent prostate cancer cells via Aurora A-mediated stabilization of SOD2

doi: 10.1016/j.isci.2026.115739

Figure Lengend Snippet: Aurora A overexpression suppresses SOD2 expression and attenuates GUTK-induced apoptosis in reactivating quiescent PCa cells (A and B) RT-PCR validation of Aurora A mRNA overexpression in LNCaP (A) and DU145 (B) cells stably transfected with either empty vector (EV) or an Aurora A expression construct (Aurora A OE). (C and D) Immunoblot analysis of Aurora A and SOD2 protein levels in quiescent EV control or Aurora A OE LNCaP (C) and DU145 (D) cells at the indicated times after cell cycle re-entry. GAPDH served as the loading control. Quantification of relative protein levels is shown in the right panel. (E and F) Apoptosis analysis by Annexin V-FITC/PI staining and flow cytometry in EV control or Aurora A OE LNCaP (E) and DU145 (F) cells treated with or without 20 μM GUTK for the indicated times during cell cycle re-entry. Quantification of apoptotic cell percentages is shown in the right panel. (G and H) ΔΨm analysis by JC-1 staining and flow cytometry in quiescent EV control and Aurora A OE LNCaP (G) and DU145 (H) cells treated with or without 20 μM GUTK for the indicated times during cell cycle re-entry. Quantification of cells with low ΔΨm is shown in the right panel. ΔΨm: mitochondrial membrane potential. Data are presented as mean ± SD from three independent experiments. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus indicated group.

Article Snippet: Sections were subjected to hematoxylin and eosin (H&E) staining or immunohistochemical staining using primary antibodies against Ki-67 (ab16667, Abcam, Shanghai, China), SOD2 (ET1701-54, HUABIO, Hangzhou, China), Cleaved Caspase 3 (#9661, Cell Signaling Technology, Danvers, USA) and Aurora A (ET1609-22, HUABIO, Hangzhou, China).

Techniques: Over Expression, Expressing, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Stable Transfection, Transfection, Plasmid Preparation, Construct, Western Blot, Control, Staining, Flow Cytometry, Membrane

Journal: iScience

Article Title: GUTK induces apoptosis in reactivating quiescent prostate cancer cells via Aurora A-mediated stabilization of SOD2

doi: 10.1016/j.isci.2026.115739

Figure Lengend Snippet: GUTK combined with docetaxel enhances the reduction of orthotopic prostate tumor growth in C57BL/6 mice (A) Experimental schematic of the orthotopic prostate cancer model and treatment schedule in C57BL/6 mice established using RM-1 murine PCa cells. Mice were randomized into four groups: vehicle, GUTK, docetaxel, or combination treatment ( n = 6 per group). (B) Representative images of major organ morphology (heart, liver, spleen, lungs, kidneys) collected on day 9. (C) Daily body weight measurements throughout the experiment period. (D) Representative images of dissected tumors collected at the day 9 endpoint. (E) Tumor volume quantification at the endpoint. (F) Representative immunohistochemical staining of tumor sections for H&E, Ki-67, SOD2, Aurora A and cleaved caspase 3. Scale bars, 50 μm. Quantification of relative protein levels is shown in the lower panel. Solvent A represents Tween 80: ethanol: saline = 20: 13: 67. Solvent B represents Cremophor EL: Ethanol: 5% glucose = 1: 1: 38. Data are presented as mean ± SEM for each group. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus vehicle control group.

Article Snippet: Sections were subjected to hematoxylin and eosin (H&E) staining or immunohistochemical staining using primary antibodies against Ki-67 (ab16667, Abcam, Shanghai, China), SOD2 (ET1701-54, HUABIO, Hangzhou, China), Cleaved Caspase 3 (#9661, Cell Signaling Technology, Danvers, USA) and Aurora A (ET1609-22, HUABIO, Hangzhou, China).

Techniques: Immunohistochemical staining, Staining, Solvent, Saline, Control

Journal: Journal of Cellular Physiology

Article Title: Maternal Obesity Programs Adipogenic Commitment in Neonatal Mesenchymal Stem Cells: A Link to Redox‐Dependent FOXO1 Signaling

doi: 10.1002/jcp.70178

Figure Lengend Snippet: Induced ROS levels in NW‐MSCs and OB‐MSCs. (A) Top: MSCs were incubated with H2DCFDA after incubation with or without H₂O₂ (400 μM) or tBHP (100 μM) to assess induced ROS levels. Two‐way ANOVA: Effect of H₂O₂ ( p = 0.0001), tBHP ( p < 0.0001) and obesity ( p = 0.01) (median ± range, n = 6). Middle: MSCs were incubated with DHE after incubation with or without Antimycin A (10 μM) to assess O 2 •‐ . Two‐way ANOVA: Effect of Antimycin A ( p < 0.0001) and obesity ( p = 0.9) (median ± range, n = 6). Bottom : MSCs were incubated with mitoSOX after incubation with or without Antimycin A (10 μM) to assess mitochondrial O 2 •‐ . Two‐way ANOVA: Effect of Antimycin A ( p = 0.002) and obesity ( p = 0.4) (median ± range, n = 6). For all graphs, Tukey's range post hoc test comparisons vs basal condition: # p < 0.05; ### p < 0.001; #### p < 0.0001). (B) SOD1/2, GPX1 and CAT gene expression were measured for basal and H₂O₂‐induced conditions (250 μM) (median ± range, n = 7 NW‐MSCs and 7 OB‐MSCs). Two‐way ANOVA: Effect of BMI (SOD1: p = 0.001; SOD2: p < 0.0001; GPX1: p < 0.0001; CAT: p = 0.003. * p < 0.05 Tukey's range post hoc test NW‐MSCs versus OB‐MSCs. # p < 0.05 Tukey´s range post hoc test basal vs induced.

Article Snippet: SOD2, a manganese‐dependent SOD (MnSOD), is the primary enzyme responsible for scavenging O 2 •‐ in the inner mitochondrial matrix and represents the first line of defense against mitochondrial oxidative damage (Hewitt and Degnan ).

Techniques: Incubation, Gene Expression