Journal: IBRO Neuroscience Reports

Article Title: TNKS1 mediates the PTEN-PI3K/AKT pathway to regulate glycolysis and proliferation in gliomas

doi: 10.1016/j.ibneur.2026.01.007

Figure Lengend Snippet: TNKS1 can activate the PI3K/AKT pathway by inhibiting the expression of PTEN in glioma cells.Western blotting was used to detect the expression of PTEN in U87 and U251 cell lines (A); the CCK-8 assay was used to measure the viability of U87 and U251 glioma cells treated with different concentrations of LY294002 and 740 Y-P (B); Western blotting was also used to detect the expression of PI3K, p-AKT/AKT in U87 and U251 cell lines (C).*P < 0.05 vs si-NC，@P < 0.05 vs si-NC+Agonist，#P < 0.05 vs si-TNKS1，&P < 0.05 vs OE-TNKS1.

Article Snippet: The cells were then cultured in the incubator for another 12 h. Subsequently, the cells in each well were randomly divided into groups: the si-NC+agonist group, the si-TNKS1 +agonist group, and the OE-TNKS1 +inhibitor group were treated with PI3K agonist 740 Y-P (HY-P0175, MCE, USA) and PI3K inhibitor LY294002 (HY-10108, MCE, USA) and continued to be cultured for 24 h. The TNKS1-siRNA empty vector group (si-NC), the TNKS1-siRNA group (si-TNKS1), and the TNKS1 overexpression group (OE-TNKS1) were treated with an equivalent amount of diluent and continued to be cultured for 24 h.

Techniques: Expressing, Western Blot, CCK-8 Assay

Journal: IBRO Neuroscience Reports

Article Title: TNKS1 mediates the PTEN-PI3K/AKT pathway to regulate glycolysis and proliferation in gliomas

doi: 10.1016/j.ibneur.2026.01.007

Figure Lengend Snippet: TNKS1 can regulate the proliferation and glycolysis of glioma cells by mediating the PTEN-PI3K/AKT pathway.Cell viability of various groups in U87 and U251 cell lines (A); detection of glucose (GLU) and lactate (LA) levels in the culture supernatant of various groups in U87 and U251 cell lines using biochemical assay kits (B); qPCR was used to detect the mRNA levels of GLUT1 and HK2 in various groups of U87 and U251 cell lines (C). Statistical significance *p < 0.05, **p < 0.01, ***p < 0.001, **** p < 0.0001,"ns" P ≥ 0.05.

Article Snippet: The cells were then cultured in the incubator for another 12 h. Subsequently, the cells in each well were randomly divided into groups: the si-NC+agonist group, the si-TNKS1 +agonist group, and the OE-TNKS1 +inhibitor group were treated with PI3K agonist 740 Y-P (HY-P0175, MCE, USA) and PI3K inhibitor LY294002 (HY-10108, MCE, USA) and continued to be cultured for 24 h. The TNKS1-siRNA empty vector group (si-NC), the TNKS1-siRNA group (si-TNKS1), and the TNKS1 overexpression group (OE-TNKS1) were treated with an equivalent amount of diluent and continued to be cultured for 24 h.

Techniques:

Journal: IBRO Neuroscience Reports

Article Title: TNKS1 mediates the PTEN-PI3K/AKT pathway to regulate glycolysis and proliferation in gliomas

doi: 10.1016/j.ibneur.2026.01.007

Figure Lengend Snippet: TNKS1 can regulate the PTEN-PI3K/AKT pathway in glioma xenografts.Western blotting was used to detect the protein expression levels of PTEN, PI3K, p-AKT, and AKT in cells from each group.* P < 0.05 vs TNKS1-siRNA NC, @ P < 0.05 vs TNKS1-siRNA，# P < 0.05 vs Inhibitor.

Article Snippet: The cells were then cultured in the incubator for another 12 h. Subsequently, the cells in each well were randomly divided into groups: the si-NC+agonist group, the si-TNKS1 +agonist group, and the OE-TNKS1 +inhibitor group were treated with PI3K agonist 740 Y-P (HY-P0175, MCE, USA) and PI3K inhibitor LY294002 (HY-10108, MCE, USA) and continued to be cultured for 24 h. The TNKS1-siRNA empty vector group (si-NC), the TNKS1-siRNA group (si-TNKS1), and the TNKS1 overexpression group (OE-TNKS1) were treated with an equivalent amount of diluent and continued to be cultured for 24 h.

Techniques: Western Blot, Expressing

Journal: IBRO Neuroscience Reports

Article Title: TNKS1 mediates the PTEN-PI3K/AKT pathway to regulate glycolysis and proliferation in gliomas

doi: 10.1016/j.ibneur.2026.01.007

Figure Lengend Snippet: TNKS1 can activate the PI3K/AKT pathway by inhibiting the expression of PTEN in glioma cells.Western blotting was used to detect the expression of PTEN in U87 and U251 cell lines (A); the CCK-8 assay was used to measure the viability of U87 and U251 glioma cells treated with different concentrations of LY294002 and 740 Y-P (B); Western blotting was also used to detect the expression of PI3K, p-AKT/AKT in U87 and U251 cell lines (C).*P < 0.05 vs si-NC，@P < 0.05 vs si-NC+Agonist，#P < 0.05 vs si-TNKS1，&P < 0.05 vs OE-TNKS1.

Article Snippet: The inhibitor group received intraperitoneal injections of LY294002 (HY-10108, MCE, USA) at a dose of 15 mg/kg, once daily for one week.

Techniques: Expressing, Western Blot, CCK-8 Assay

Journal: iScience

Article Title: Thrombospondin 1 aggravates cardiac remodeling in heart failure with preserved ejection fraction by inhibiting mitophagy

doi: 10.1016/j.isci.2026.114639

Figure Lengend Snippet: Thbs1 knockdown alleviates cardiomyocyte hypertrophy, oxidative stress, mitochondrial dysfunction, and mitophagy impairment in vitro (A) Schematic diagram of the in vitro pathological stimulation protocol. H9c2 cells were treated with isoproterenol (ISO) and macrophage-conditioned medium (MCM) derived from LPS+ATP-activated macrophages, with or without Thbs1 knockdown. (B and C) mRNA levels of hypertrophic markers ANP (B) and BNP (C) under pathological stimulation ( n = 3 biological replicates per group, each measured in triplicate). (D) Representative fluorescence images of mitochondrial reactive oxygen species (ROS) detected using MitoSOX Red. Scale bars, 20 μm. (E) Quantification of MitoSOX Red fluorescence intensity ( n = 6 biological replicates per group, each measured in triplicate). (F) JC-1 staining to assess mitochondrial membrane potential (MMP), representative images shown. Scale bars, 20 μm. (G) Quantification of JC-1 aggregates/monomers ratio ( n = 6 biological replicates per group, each measured in triplicate). (H) Immunofluorescence images showing LC3 (red) and VDAC (green) colocalization; nuclei were stained with DAPI (blue). Scale bars, 20 μm. (I) Representative western blot images of mitophagy-related proteins LC3, p62, PINK1, and Parkin. (J–M) Quantification of protein expression: LC3-II/I (J), p62 (K), PINK1 (L), and Parkin (M) ( n = 3 biological replicates per group, each measured in duplicate). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, indicating the level of statistical significance for each comparison.

Article Snippet: Pharmacologic modulation of the PI3K/Akt/mTOR pathway was achieved by pretreating H9c2 cells with 10 μM LY294002 (PI3K inhibitor; HY-10108, MedChemExpress) or 10 μM SC79 (an Akt activator; HY-18749, MedChemExpress) for 1 h before stimulation.

Techniques: Knockdown, In Vitro, Derivative Assay, Fluorescence, Staining, Membrane, Immunofluorescence, Western Blot, Expressing, Comparison

Journal: iScience

Article Title: Thrombospondin 1 aggravates cardiac remodeling in heart failure with preserved ejection fraction by inhibiting mitophagy

doi: 10.1016/j.isci.2026.114639

Figure Lengend Snippet: PI3K/Akt/mTOR pathway activation reverses the protective effects of Thbs1 knockdown in H9c2 cells (A) Representative western blot images showing phosphorylation and total levels of Akt and mTOR in H9c2 cells transfected with si- Thbs1 and treated with ISO plus MCM or the Akt activator SC79. (B and C) Quantification of protein phosphorylation: p -Akt/Akt (B) and p -mTOR/mTOR (C) ( n = 3 biological replicates per group, each measured in duplicate). (D and E) Relative mRNA expression of ANP (D) and BNP (E) assessed by RT-qPCR ( n = 3 biological replicates per group, each measured in triplicate). (F and G) Intracellular ROS production measured using MitoSOX Red mitochondrial superoxide indicator; representative fluorescence images are shown (F) and quantified as mean fluorescence intensity (G) ( n = 6 biological replicates per group, each measured in triplicate). Scale bars, 20 μm. (H) Representative confocal images of mitochondria-lysosome colocalization using Mito-Tracker (green) and Lyso-Tracker (red). Scale bars, 20 μm. (I) Representative western blot images of mitophagy-related proteins LC3, p62, PINK1, and Parkin. (J–M) Quantification of mitophagy-related protein expression: LC3-II/I (J), p62 (K), PINK1 (L), and Parkin (M) ( n = 3 biological replicates per group, each measured in duplicate). Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, indicating the level of statistical significance for each comparison.

Article Snippet: Pharmacologic modulation of the PI3K/Akt/mTOR pathway was achieved by pretreating H9c2 cells with 10 μM LY294002 (PI3K inhibitor; HY-10108, MedChemExpress) or 10 μM SC79 (an Akt activator; HY-18749, MedChemExpress) for 1 h before stimulation.

Techniques: Activation Assay, Knockdown, Western Blot, Phospho-proteomics, Transfection, Expressing, Quantitative RT-PCR, Fluorescence, Comparison

Journal: iScience

Article Title: Thrombospondin 1 aggravates cardiac remodeling in heart failure with preserved ejection fraction by inhibiting mitophagy

doi: 10.1016/j.isci.2026.114639

Figure Lengend Snippet: Inhibition of PI3K/Akt/mTOR reverses effects of Thbs1 overexpression and Thbs1 binds ITGB1 in H9c2 cells (A) Representative western blot images showing phosphorylation and total levels of PI3K, Akt, and mTOR in H9c2 cells transfected with Thbs1 overexpression plasmid and treated with ISO plus MCM or the PI3K/Akt/mTOR inhibitor LY294002. (B–D) Quantification of protein phosphorylation: p-PI3K/PI3K (B), p -Akt/Akt (C), and p -mTOR/mTOR (D) ( n = 3 biological replicates per group, each measured in duplicate). (E and F) Intracellular ROS production measured using MitoSOX Red; representative fluorescence images are shown (E) and quantified as mean fluorescence intensity (F) ( n = 6 biological replicates per group, each measured in triplicate). Scale bars, 20 μm. (G) Representative confocal images showing mitochondria-lysosome colocalization using Mito-Tracker (green) and Lyso-Tracker (red). Scale bars, 20 μm. (H) Representative western blot images of mitophagy-related proteins LC3, p62, PINK1, and Parkin. (I–L) Quantification of mitophagy-related protein expression: LC3-II/I (I), p62 (J), PINK1 (K), and Parkin (L) ( n = 3 biological replicates per group, each measured in duplicate). (M) Co-immunoprecipitation (Co/IP) assays showing the interaction between Thbs1 and ITGB1 in H9c2 cells. Data are presented as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, indicating the level of statistical significance for each comparison.

Article Snippet: Pharmacologic modulation of the PI3K/Akt/mTOR pathway was achieved by pretreating H9c2 cells with 10 μM LY294002 (PI3K inhibitor; HY-10108, MedChemExpress) or 10 μM SC79 (an Akt activator; HY-18749, MedChemExpress) for 1 h before stimulation.

Techniques: Inhibition, Over Expression, Western Blot, Phospho-proteomics, Transfection, Plasmid Preparation, Fluorescence, Expressing, Immunoprecipitation, Co-Immunoprecipitation Assay, Comparison