Journal: Nature Communications

Article Title: Podocyte GSK3 is an evolutionarily conserved critical regulator of kidney function

doi: 10.1038/s41467-018-08235-1

Figure Lengend Snippet: Hippo signalling is disrupted in GSK3 deficient podocytes in vitro. a Summary of proteomics results for Ajuba in Cre treated wild-type (CreWT n = 3), Cre treated floxed GSK3α/β (GSK3KO n = 4) and non-Cre treated floxed GSK3α/β (GSK3FL n = 3) podocytes. ANOVA p = 0.004, Tukey post hoc analysis ** p < 0.01 *** p < 0.001. b Representative western blot of analysis of ciGSK3DKO cells shows increased expression of Ajuba relative to controls, n = 3–4. See also Supplementary Fig. . c , d Representative western blots of wild-type mouse podocytes incubated with 20 mM LiCl ( c ) and 3 μM CHIR99201 ( d ) showing increased expression of ajuba and Cyclin B1, n = 3. See also Supplementary Figs. and c. e – g Immunofluorescence analysis showing increased nuclear YAP/TAZ staining in ciGSK3DKO cells 24 h after induction of gene knockout ( e ) n = 3, wild-type podocytes after 24 h 20 mM LiCl ( f ) n = 4 and 24 h 3 μM CHIR99201 ( g ) n = 8. Unpaired t test * p < 0.05. Data are presented as the mean ± SEM

Article Snippet: Fig. 6 Hippo signalling is disrupted in GSK3 deficient podocytes in vitro. a Summary of proteomics results for Ajuba in Cre treated wild-type (CreWT n = 3), Cre treated floxed GSK3α/β (GSK3KO n = 4) and non-Cre treated floxed GSK3α/β (GSK3FL n = 3) podocytes.

Techniques: In Vitro, Western Blot, Expressing, Incubation, Immunofluorescence, Staining, Gene Knockout

Journal: Nature Communications

Article Title: Podocyte GSK3 is an evolutionarily conserved critical regulator of kidney function

doi: 10.1038/s41467-018-08235-1

Figure Lengend Snippet: Hippo signalling is disrupted in GSK3 deficient podocytes in vivo. a , b Western blot ( a ) and quantification ( b ) of glomeruli isolated from mpodGSK3DKO and littermate control mice showing increased expression of Ajuba, n = 2 mice per group. c , d Immunohistochemistry showing increased Ajuba (top panels) and YAP/TAZ nuclear translocation (arrowed, bottom panels) in the podocytes of podGSK3DKO ( c ) and mpodGSK3DKO ( d ) mice. Representative immunohistochemistry and quantification shown (>5 glomeruli per mouse analysed, 3 mice per group, ANOVA *** p < 0.001. Scale bar = 25 μm). e Immunohistochemistry showing an increase in the number of cells positive for the YAP/TAZ TEAD target Cyclin A2. Three mice from each group analysed, t test * p < 0.5. f Immunofluorescence staining showing increased expression of c-myc in podGSK3DKO and mpodGSK3DKO mice. Three mice analysed per group, t test *** p < 0.001. Scale bar = 50 μm. g Representative immunohistochemistry using an anti-Ajuba antibody in a glomerulus from a rat given high dose lithium for 6 months and in a biopsy from a patient on long-term lithium therapy. Ajuba expression is increased with lithium treatment relative to control tissue. Scale bar = 25 μm. Data are presented as the mean ± SEM

Article Snippet: Fig. 6 Hippo signalling is disrupted in GSK3 deficient podocytes in vitro. a Summary of proteomics results for Ajuba in Cre treated wild-type (CreWT n = 3), Cre treated floxed GSK3α/β (GSK3KO n = 4) and non-Cre treated floxed GSK3α/β (GSK3FL n = 3) podocytes.

Techniques: In Vivo, Western Blot, Isolation, Control, Expressing, Immunohistochemistry, Translocation Assay, Immunofluorescence, Staining

Journal: Nature Communications

Article Title: Podocyte GSK3 is an evolutionarily conserved critical regulator of kidney function

doi: 10.1038/s41467-018-08235-1

Figure Lengend Snippet: Verteporfin attenuates the effects of GSK3 loss in Drosophila . a Knock down of shaggy ( sgg ) in nephrocytes prevents Duf translocation to the cell surface and is rescued by verteporfin. Flies were raised at the non-permissive temperature (18 °C, which does not allow sgg RNAi expression) and then cultured for 48 h at either the same temperature, or at the higher temperature (29 °C, which allows sgg RNAi expression). Cells were maintained in the presence or absence of 400 nM verteporfin. Nephrocytes were stained with antisera raised to the slit diaphragm protein Dumbfounded (Duf, cyan) as well as wheatgerm agglutinin (magenta). Duf was expressed at the surface of nephrocytes when RNAi was prevented but surface expression was completely prevented in when sgg RNAi was initiated. The lack of surface expression was rescued by the presence of verteporfin. Scale bar = 50 µm. b High magnification images showing rescue of surface expression by verteporfin Sale bar = 40 µm. c Knock down of shaggy ( sgg ) in nephrocytes prevents Duf translocation to the cell surface and is rescued by verteporfin. n = 6–7 flies per genotype. ANOVA, ** p < 0.01 for sgg TARGET at 29 °C having fewer Duf + ve nephrocytes than other groups. d Verteporfin rescues lithium induced toxicity in nephrocytes. Adult wild-type flies were reared to between 3–7 days of age, dissected to reveal the heart and nephrocytes and cultured for 24 h in the presence or absence of lithium (20 mM), verteporfin (400 nM) or both. Scale bar top panel = 30 μm; bottom panel = 10 μm. e Nephrocyte viability was assessed with the vital stain calcein-AM, with nephrocytes being visualised using wheatgerm agglutinin. Lithium had a significant impact on nephrocyte viability (**** p < 0.0001) and this was rescued by the addition of verteporfin (**** p < 0.0001). Means compared by one-way ANOVA followed by Tukey’s HSD. n = 12 flies from two independent experiments. f Cartoon showing the effect of podocyte GSK3 loss on hippo signalling. GSK3 deletion increases the expression of Ajuba, preventing the phosphorylation of YAP/TAZ and allowing its translocation into the nucleus. This results in cell cycle re-entry, mitotic catastrophe and ultimately apoptosis. Data are presented as the mean ± SEM

Article Snippet: Fig. 6 Hippo signalling is disrupted in GSK3 deficient podocytes in vitro. a Summary of proteomics results for Ajuba in Cre treated wild-type (CreWT n = 3), Cre treated floxed GSK3α/β (GSK3KO n = 4) and non-Cre treated floxed GSK3α/β (GSK3FL n = 3) podocytes.

Techniques: Knockdown, Translocation Assay, Expressing, Cell Culture, Staining, Phospho-proteomics

Journal: PLoS ONE

Article Title: Type 2 diabetes is associated with loss of HDL endothelium protective functions

doi: 10.1371/journal.pone.0192616

Figure Lengend Snippet: Protein quantified in the targeted proteomics analysis.

Article Snippet: Proteomics data are available from figshare (DOI: 10.6084/m9.figshare.5890555 ), as well as the PanoramaWeb public repository for targeted proteomics results ( https://panoramaweb.org/labkey/VaisarHDL_T2D.url ).

Techniques: Targeted Proteomics, Binding Assay

Journal: Cell Death & Disease

Article Title: A Lamp2a-linked RNA secreted by ADSCs prevents ENO1–lactylation–glycolysis feedback and cell malignant behavior in triple-negative breast cancer

doi: 10.1038/s41419-026-08517-3

Figure Lengend Snippet: A Immunofluorescence analysis of HIF-1α and PKM2 expression in normal mammary tissue, ER+ BC (representing non-TNBC tumors), and TNBC tissues. TNBC shows significantly higher expression of both glycolytic regulators compared to ER+ BC and normal tissue. Scale bar: 50 μm. B Heatmap of significantly differentially expressed proteins (DEPs) between TNBC and ER+ BC, highlighting metabolic alterations. C Gene Ontology (GO) enrichment analysis of upregulated DEPs in TNBC, revealing strong associations with glycolysis, gluconeogenesis, and pyruvate metabolism. D Western blot validation of key glycolytic enzymes (HK1, GPI, PFKP, PGK1, ENO1, PKM2, HIF-1α) in TNBC versus ER+ BC. β-actin serves as a loading control. E Principal component analysis (PCA) plot of 633 breast tumor samples from integrated GSE datasets after batch-effect correction. F Venn diagram illustrating the number of commonly upregulated and downregulated genes in TNBC across multiple datasets (GSE, TCGA-BRCA, METABRIC). G GO-BP enrichment analyses of integrated datasets ( GSE65194 , GSE103091 , GSE76275 , TCGA-BRCA, METABRIC) in TNBC versus non-TNBC, demonstrating significant enrichment of glycolysis-related gene sets (FDR < 0.05). H , I the relationship between patient survival time and DEGs using a Cox proportional hazards model and clinical survival and prognosis information provided in the GSE103091 dataset and METABRIC database. J , K OCR and ECAR were tested in TNBC cell line MDA-MB-231, the Luminal cell line MCF-7, the HER2-positive cell line SKBR3, and the non-tumorigenic human mammary epithelial cell line MCF-10A. Data represent mean ± SEM ( n = 3 for Proteomics and Western blot assays; n = 5 for OCR and ECAR assays); * p < 0.05, ** p < 0.01, *** p < 0.001 (unpaired Student’s t test for comparisons involving two treatment groups, or a one-way ANOVA for comparisons involving more than two groups).

Article Snippet: Proteomics were conducted with the help of Genechem Co., Ltd. (Shanghai, China).

Techniques: Immunofluorescence, Expressing, Western Blot, Biomarker Discovery, Control