Journal: Renal Failure

Article Title: Canagliflozin ameliorates high-salt-induced renal injury and premature aging in male Dahl salt-sensitive rats, with associated changes in SIRT6/HIF-1α signaling

doi: 10.1080/0886022X.2025.2546624

Figure Lengend Snippet: Representative images of senescence-associated β-galactosidase (SA-β-gal) staining and the expression of aging-related proteins in the kidneys of the three groups of rats. A. Renal SA-β-gal staining. Magnification, ×200; scale bar, 50 μm. B. Expression of the aging-related proteins P53 and P21 in the kidneys. C. SA-β-gal-positive area in rat kidneys ( n = 4). D. P53 protein expression. E. P21 protein expression. (D, E, n = 6). NSD: normal salt dahl salt-sensitive rats; HSD: high salt dahl salt-sensitive rats; HSD + CANA: high salt dahl salt-sensitive rats + canagliflozin (30 mg/kg/day). HSD vs. NSD: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. HSD vs. HSD + CANA: # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001.

Article Snippet: The antibodies used for rat tissues included rabbit monoclonal anti-SIRT6 (1:1000, Abcam, ab191385), rabbit polyclonal anti-p53 (1:800, Proteintech, 10442-1-AP), rabbit polyclonal anti-p21 (1:5000, Proteintech, 10355-1-AP), rabbit polyclonal anti-HIF-1α (1:100, affinity, AF1009), and rabbit monoclonal anti-αSMA (alpha smooth muscle, 1:1000, Zenbio, R380653) antibodies.

Techniques: Staining, Expressing

Journal: Renal Failure

Article Title: Canagliflozin ameliorates high-salt-induced renal injury and premature aging in male Dahl salt-sensitive rats, with associated changes in SIRT6/HIF-1α signaling

doi: 10.1080/0886022X.2025.2546624

Figure Lengend Snippet: Immunohistochemical results of the three groups of rats. A. Representative image of SIRT6 in the kidney. B. Representative image of HIF-1α in the kidney. C. Representative image of α-SMA in the kidney. D. Representative image of P53 in the kidney. E. Representative image of P21 in the kidney. Magnification, ×200; scale bar, 20 μm. NSD: normal salt dahl salt-sensitive rats; HSD: high salt dahl salt-sensitive rats; HSD + CANA: high salt dahl salt-sensitive rats + canagliflozin (30 mg/kg/day). HSD vs. NSD: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. HSD vs. HSD + CANA: # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 ( n = 6).

Article Snippet: The antibodies used for rat tissues included rabbit monoclonal anti-SIRT6 (1:1000, Abcam, ab191385), rabbit polyclonal anti-p53 (1:800, Proteintech, 10442-1-AP), rabbit polyclonal anti-p21 (1:5000, Proteintech, 10355-1-AP), rabbit polyclonal anti-HIF-1α (1:100, affinity, AF1009), and rabbit monoclonal anti-αSMA (alpha smooth muscle, 1:1000, Zenbio, R380653) antibodies.

Techniques: Immunohistochemical staining

Journal: Molecular Medicine Reports

Article Title: CDK5 targets p21 CIP1 to regulate thyroid cancer cell proliferation and malignancy in patients

doi: 10.3892/mmr.2025.13547

Figure Lengend Snippet: CDK5 interacts with p21. (A) Structural prediction of the interaction between CDK5 and p21 using AlphaFold 3.0. CDK5 is shown in red and p21 in green. (B) Detailed visualization of the CDK5-p21 interaction interface using UCSF Chimera software. CDK5 is shown in surface representation, while p21 is depicted in a ribbon diagram. The binding sites are highlighted. Heat map of the interaction interface indicating the degree of interaction strength between CDK5 and p21, with red representing strong interaction sites and blue representing weak or no interaction. (C) Sequence alignment of CDK5 and p21 showing the interacting regions predicted by AlphaFold. Residues involved in the interaction are highlighted in red for CDK5 and green for p21. (D) Immunoprecipitation assay demonstrating the interaction between CDK5 and p21 in TPC-1 cells. Cells were treated with 10 µM MG132 to inhibit proteasomal degradation. CDK5 was immunoprecipitated from cell lysates and co-precipitated p21 was detected by western blotting. Input lysates (bottom panels) show the expression levels of p21, CDK5 and β-actin, the latter was used as the loading control. The results confirm the interaction between CDK5 and p21 and suggested that CDK5 regulates p21 stability via the ubiquitin-proteasome pathway. CDK5, cyclin-dependent kinase 5.

Article Snippet: The tissue sections were incubated overnight at 4°C with the following primary antibodies: Mouse monoclonal anti-CDK5 (cat. no. sc-249; Santa Cruz Biotechnology, Inc.; 1:20) or rabbit polyclonal anti-p21 (cat. no. 2947; Cell Signaling Technology, Inc.; 1:50).

Techniques: Structural Proteomics, Software, Binding Assay, Sequencing, Immunoprecipitation, Western Blot, Expressing, Control, Ubiquitin Proteomics

Journal: Molecular Medicine Reports

Article Title: CDK5 targets p21 CIP1 to regulate thyroid cancer cell proliferation and malignancy in patients

doi: 10.3892/mmr.2025.13547

Figure Lengend Snippet: CDK5 downregulates p21 in TC cells. (A) Western blot analysis of p21 protein expression levels in BCPAP cells transfected with EV or a CDK5 overexpression plasmid. Cells were treated with 50 µg/ml CHX for 0, 2, 4 or 6 h to inhibit protein synthesis. Short and long exposures of p21 are shown. (B) Quantification of p21 protein levels. Data are presented as the fold change relative to time 0 for EV and CDK5 transfected cells. (C) Western blot analysis of p21 and CDK5 protein expression levels in BCPAP cells transfected with EV or CDK5, with or without treatment with 10 µM MG132 for 6 h. (D) Quantification of p21 protein expression. Data are presented as fold change relative to EV-transfected mock-treated cells. (E) Immunofluorescence staining of p21 (green) in BCPAP cells transfected with vector or CDK5, with or without MG132 treatment. The results indicate that CDK5 overexpression reduces p21 levels in TC cells and this effect was reversed following proteasome inhibition. Scale bar, 20 µm. (F) Co-immunoprecipitation analysis of CDK5 and p21 was performed to investigate the interaction between CDK5 and p21 in thyroid cancer cells. Panels show the interaction of CDK5 with WT p21 and MT p21 following proteasome inhibition using MG132. Red arrows indicate the presence of p21 in the immunoprecipitated complex. (G) Cell counting assay was performed to evaluate p21 WT and MT p21 on cell proliferation. For all blots, β-actin was used as the loading control. Values are presented as the mean ± SD from three independent experiments. Data were compared using an unpaired Student's t-test. *P<0.05, **P<0.01, ***P<0.001. CDK5, cyclin-dependent kinase; 5TC, thyroid cancer; EV, empty vector; HCX, cycloheximide; n.s., not significant; WT, wild-type; MT, S130A mutant.

Article Snippet: The tissue sections were incubated overnight at 4°C with the following primary antibodies: Mouse monoclonal anti-CDK5 (cat. no. sc-249; Santa Cruz Biotechnology, Inc.; 1:20) or rabbit polyclonal anti-p21 (cat. no. 2947; Cell Signaling Technology, Inc.; 1:50).

Techniques: Western Blot, Expressing, Transfection, Over Expression, Plasmid Preparation, Immunofluorescence, Staining, Inhibition, Immunoprecipitation, Cell Counting, Control, Mutagenesis

Journal: Molecular Medicine Reports

Article Title: CDK5 targets p21 CIP1 to regulate thyroid cancer cell proliferation and malignancy in patients

doi: 10.3892/mmr.2025.13547

Figure Lengend Snippet: CDK5 negatively regulates p21 in TC cells. Reverse transcription-quantitative PCR analysis of (A) CDK5 and (B) CDKN1A mRNA expression in TC cell lines BCPAP and TPC-1. (C) Western blot analysis of p21 and CDK5 protein levels in BCPAP and TPC-1 cells treated with or without 10 µM MG132 for 6 h. (D) Western blot analysis of p21 and CDK5 protein levels in TPC-1 and BCPAP cells transfected with shGFP or shCDK5 (#1, #2, and #3). (E) Quantitative analysis of CDK5 mRNA levels in both TPC-1 and BCPAP cells. Data are presented as the fold change relative to shGFP-transduced cells. The results demonstrated that knockdown of CDK5 increases p21 protein expression levels in TC cells. Values are presented as the mean ± SD from three independent experiments. Data were compared using an unpaired Student's t-test. *P<0.05, **P<0.01, ***P<0.001 vs. shGFP. CDK5, cyclin-dependent kinase; TC, papillary thyroid cancer; sh, short hairpin; n.s., not significant.

Article Snippet: The tissue sections were incubated overnight at 4°C with the following primary antibodies: Mouse monoclonal anti-CDK5 (cat. no. sc-249; Santa Cruz Biotechnology, Inc.; 1:20) or rabbit polyclonal anti-p21 (cat. no. 2947; Cell Signaling Technology, Inc.; 1:50).

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Transfection, Knockdown

Journal: Molecular Medicine Reports

Article Title: CDK5 targets p21 CIP1 to regulate thyroid cancer cell proliferation and malignancy in patients

doi: 10.3892/mmr.2025.13547

Figure Lengend Snippet: Analysis of TCGA data showing the clinical relevance of CDK5 and p21 in thyroid cancer. (A) Scatter plot showing the correlation between CDK5 and p21 mRNA expression in thyroid cancer samples from TCGA. Pearson correlation coefficient (r) and P-value are indicated. (B) Kaplan-Meier overall survival analysis of patients with thyroid cancer stratified by high and low CDK5 expression levels. (C) Kaplan-Meier overall survival analysis of patients with thyroid cancer stratified by high and low p21 expression levels. Survival was compared using a log-rank test. (D) Pathological stage plot showing CDK5 expression levels across different stages of thyroid cancer. (E) Pathological stage plot illustrating p21 expression levels across different stages of thyroid cancer. Pathological stage plots were obtained from GEPIA, and differential gene expression was analyzed by one-way ANOVA ( https://gepia.cancer-pku.cn/ ). Higher CDK5 expression was associated with poorer overall survival, while p21 expression was not markedly associated with survival outcomes. TCGA, The Cancer Genome Atlas; CDK5, cyclin-dependent kinase.

Article Snippet: The tissue sections were incubated overnight at 4°C with the following primary antibodies: Mouse monoclonal anti-CDK5 (cat. no. sc-249; Santa Cruz Biotechnology, Inc.; 1:20) or rabbit polyclonal anti-p21 (cat. no. 2947; Cell Signaling Technology, Inc.; 1:50).

Techniques: Expressing, Gene Expression

Journal: Molecular Medicine Reports

Article Title: CDK5 targets p21 CIP1 to regulate thyroid cancer cell proliferation and malignancy in patients

doi: 10.3892/mmr.2025.13547

Figure Lengend Snippet: IHC analysis of CDK5 and p21 nuclear expression in papillary thyroid cancer patient specimens. (A) IHC staining for CDK5 and p21 in patients with thyroid cancer demonstrating the relationship between CDK5 and p21 nuclear expression. Left panels, CDK5 staining showed strong nuclear localization in several patient samples, particularly in tumor cells. The nuclear expression of CDK5 was more prominent in aggressive regions of the thyroid cancer samples, suggesting a role in tumor proliferation. Right panels, p21 staining was inversely correlated with CDK5. In samples where CDK5 was strongly expressed in the nucleus, p21 nuclear staining was weak or absent. Conversely, in areas were CDK5 expression was low, p21 nuclear localization was more evident. This expression pattern supported the hypothesis that CDK5 negatively regulated p21 expression at the nuclear level, contributing to cancer cell cycle dysregulation and malignancy. Scale bar, 50 µm. (B) The heatmap illustrates the correlation between nuclear CDK5 and p21 expression across multiple samples, with darker red indicating higher CDK5 expression and lower p21 levels. (C) Significant differences in expression patterns between groups with high CDK5/low p21 and low CDK5/high p21 expression (*P<0.05). This suggested that elevated nuclear CDK5 was associated with decreased p21 expression, supporting the hypothesis that CDK5 promoted thyroid cancer progression. IHC, immunohistochemistry; CDK5, cyclin-dependent kinase.

Article Snippet: The tissue sections were incubated overnight at 4°C with the following primary antibodies: Mouse monoclonal anti-CDK5 (cat. no. sc-249; Santa Cruz Biotechnology, Inc.; 1:20) or rabbit polyclonal anti-p21 (cat. no. 2947; Cell Signaling Technology, Inc.; 1:50).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Molecular Medicine Reports

Article Title: CDK5 targets p21 CIP1 to regulate thyroid cancer cell proliferation and malignancy in patients

doi: 10.3892/mmr.2025.13547

Figure Lengend Snippet: Graphical abstract. The present study examined the interaction between CDK5 and the cyclin-dependent kinase inhibitor p21 CIP1 . CDK5 promoted the degradation of p21 through the ubiquitin-mediated pathways, thereby reducing the tumor-suppressive effects of p21. High CDK5 levels were associated with increased tumor malignancy and worse survival outcomes, while higher p21 expression was associated with an improved prognosis. TC stages and aggressive tumors exhibit elevated CDK5 and reduced p21 levels. This suggested that CDK5-mediated degradation of p21 contributed to TC progression. CDK5, cyclin-dependent kinase 5; TC, thyroid cancer.

Article Snippet: The tissue sections were incubated overnight at 4°C with the following primary antibodies: Mouse monoclonal anti-CDK5 (cat. no. sc-249; Santa Cruz Biotechnology, Inc.; 1:20) or rabbit polyclonal anti-p21 (cat. no. 2947; Cell Signaling Technology, Inc.; 1:50).

Techniques: Ubiquitin Proteomics, Expressing

Journal: Frontiers in Pharmacology

Article Title: Exploring the potential anti-senescence effects of soybean-derived peptide Soymetide in mice hippocampal neurons via the Wnt/β-catenin pathway

doi: 10.3389/fphar.2025.1510337

Figure Lengend Snippet: Soymetide prevents doxorubicin-induced senescence in the hippocampus of mice . Hippocampal tissues were isolated from vehicle (V), doxorubicin (DOX), Soymetide (Soym)+DOX and senolytic combination (Seno)+DOX-treated mice. (A–F) Representative Western blots and corresponding densitometry (relative to (Rel.) vehicle) of p53 (A, D) , p21 (B, E) , and p16 (C, F) normalized with β-actin in the hippocampal tissues. Data represent means ± Standard Error (SE) of three mice/group. ***p < 0.001, **p < 0.01 and *p < 0.05 compared to V. ### p < 0.001, ## p < 0.01 and # p < 0.05 compared to DOX.

Article Snippet: Mouse monoclonal p53 (cat no. 2524), rabbit polyclonal p21 (cat no. 64016), phospho-LRP6 (cat no. 2568), β-catenin (cat no. 9562), rabbit monoclonal NeuN (cat no. 24307), LRP6 (cat no. 3395), Dvl2 (cat no. 3224), Axin1 (cat no. 2087), GSK-3β (cat no. 12456) and TCF3/TCF7L1 (cat no. 2883) antibodies were purchased from Cell Signaling Technology.

Techniques: Isolation, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Exploring the potential anti-senescence effects of soybean-derived peptide Soymetide in mice hippocampal neurons via the Wnt/β-catenin pathway

doi: 10.3389/fphar.2025.1510337

Figure Lengend Snippet: The Soymetide-induced reduction in senescence markers and increase in NeuN levels are Wnt3a/β-catenin-dependent in the hippocampus of doxorubicin-treated mice . Hippocampal tissues and the whole brain were isolated from vehicle (V), doxorubicin (DOX), Soymetide (Soym)+DOX, Soym + DOX + rDkk1, and Soym + DOX + iCRT3-treated mice. (A–C, E) Representative Western blots and corresponding densitometry (relative to vehicle) of p53 (A) , p21 (B) , p16 (C) and NeuN (E) normalized with β-actin in the hippocampal tissues. (D) Representative photomicrographs and corresponding bar graph (relative to vehicle) of Senescence β-Galactosidase-stained hippocampus in brain sections. Scale bar: 100 µm. Data represent means ± SE of three mice/group. The quantification of the first three groups in (D) and (E) are based on the same data sets as in . ***p < 0.001 and **p < 0.01 compared to V. ### p < 0.001, ## p < 0.01 and # p < 0.05 compared to DOX. @@@ p < 0.001, @@ p < 0.01 and @ p < 0.05 compared to Soym + DOX.

Article Snippet: Mouse monoclonal p53 (cat no. 2524), rabbit polyclonal p21 (cat no. 64016), phospho-LRP6 (cat no. 2568), β-catenin (cat no. 9562), rabbit monoclonal NeuN (cat no. 24307), LRP6 (cat no. 3395), Dvl2 (cat no. 3224), Axin1 (cat no. 2087), GSK-3β (cat no. 12456) and TCF3/TCF7L1 (cat no. 2883) antibodies were purchased from Cell Signaling Technology.

Techniques: Isolation, Western Blot, Staining

Journal: Frontiers in Pharmacology

Article Title: Exploring the potential anti-senescence effects of soybean-derived peptide Soymetide in mice hippocampal neurons via the Wnt/β-catenin pathway

doi: 10.3389/fphar.2025.1510337

Figure Lengend Snippet: Schematic of the anti-senescent effect induced by Soymetide against doxorubicin treatment in hippocampal neurons and its positive impact on cognitive function . Soymetide prevents the reduction in Wnt3a and its related proteins (Fz, p-LRP-6, Dvl, and Axin1) caused by doxorubicin, and it counters the rise in GSK-3β and the reduction of β-catenin. This inhibits the increase in senescence markers such as p53, p21, p16, β- Galactosidase activity, and SASPs, as well as the decrease in neuronal survival and learning-memory functions. In summary, Soymetide exhibits anti-senescent effects, provides neuroprotection, and induces cognitive restoration by restoring the Wnt/β-catenin pathway.

Article Snippet: Mouse monoclonal p53 (cat no. 2524), rabbit polyclonal p21 (cat no. 64016), phospho-LRP6 (cat no. 2568), β-catenin (cat no. 9562), rabbit monoclonal NeuN (cat no. 24307), LRP6 (cat no. 3395), Dvl2 (cat no. 3224), Axin1 (cat no. 2087), GSK-3β (cat no. 12456) and TCF3/TCF7L1 (cat no. 2883) antibodies were purchased from Cell Signaling Technology.

Techniques: Activity Assay