Journal: International journal of biological sciences

Article Title: Treatment for type 2 diabetes and diabetic nephropathy by targeting Smad3 signaling.

doi: 10.7150/ijbs.87820

Figure Lengend Snippet: Figure 1. Prediabetic treatment of db/db mice with SIS3 from the age of 4 weeks to 12 weeks largely improves hyperglycemia, glucose tolerance, insulin resistance and renal functional injury. (A) Fasting blood glucose and area under curve (AUC). (B, C) Blood glucose levels and AUC during intraperitoneal glucose tolerance test (IPGTT) and intraperitoneal insulin tolerance test (IPITT). (D) HbA1c levels. (E) Serum creatinine. (F) Urinary albumin/creatinine ratio (UACR). Data represents the mean ± SEM for at least 6 mice per group.

Article Snippet: To determine a safe and effective dose of SIS3 for treatment of db/db mice, groups of 6-8 male db/db and normal db/m mice were intraperitoneal (i.p.) injected with solvent (DMSO) diluted SIS3 (Selleck Chemicals, Houston, TX, USA; cat# S7959) at dosages of 1.25, 2.5, 5mg/kg.bw/day for 8 weeks from the age of 4 weeks to 12 weeks.

Techniques: Functional Assay

Journal: International journal of biological sciences

Article Title: Treatment for type 2 diabetes and diabetic nephropathy by targeting Smad3 signaling.

doi: 10.7150/ijbs.87820

Figure Lengend Snippet: Figure 2. Prediabetic treatment of db/db mice with SIS3 from the age of 4 weeks to 12 weeks protects against renal fibrosis. (A) PAS and immunostaining with antibodies against collagens I and IV, fibronectin (FN), and α-SMA in the week 12 kidney of db/m and db/db mice treated with or without SIS3 at a dose of 2.5 mg/kg/day. (B-F) Semi-quantitation of glomerular matrix deposition and accumulation of collagens I, IV, fibronectin, and α-SMA+ myofibroblasts from the immunostained sections. (G-J) Western blotting and quantitative analysis of renal fibrosis including collagen I, α-SMA, and fibronectin. (K-M) Real-time PCR analysis for mRNA expression of fibronectin, collagen I, and α-SMA in the kidney of db/m or db/db mice at 12 weeks. Data represents the mean ± SEM for at least 6 mice per group. **P<0.01, ***P<0.001, ****P<0.0001 versus normal db/m mice. #P<0.05, ##P<0.01,

Article Snippet: To determine a safe and effective dose of SIS3 for treatment of db/db mice, groups of 6-8 male db/db and normal db/m mice were intraperitoneal (i.p.) injected with solvent (DMSO) diluted SIS3 (Selleck Chemicals, Houston, TX, USA; cat# S7959) at dosages of 1.25, 2.5, 5mg/kg.bw/day for 8 weeks from the age of 4 weeks to 12 weeks.

Techniques: Immunostaining, Quantitation Assay, Western Blot, Real-time Polymerase Chain Reaction, Expressing

Journal: International journal of biological sciences

Article Title: Treatment for type 2 diabetes and diabetic nephropathy by targeting Smad3 signaling.

doi: 10.7150/ijbs.87820

Figure Lengend Snippet: Figure 3. Prediabetic treatment of db/db mice with SIS3 from the age of 4 weeks to 12 weeks protects against renal inflammation. (A-C) Immunostaining and quantitative analysis of F4/80+ macrophages and expression of MCP-1 in the kidney of db/m and db/db mice at 12 weeks of age. (D-F) Real-time PCR analysis detects IL-1β, TNF-α, and MCP-1 mRNA expression in kidney of db/m and db/db mice at 12 weeks. (G-I) ELISA analysis detects serum levels of IL-1β, TNF-α and MCP-1 in db/m and db/db mice at 12 weeks. Data represents the mean ± SEM for at least 6 mice per group. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 versus normal db/m mice. #P<0.05, ##P<0.01,

Article Snippet: To determine a safe and effective dose of SIS3 for treatment of db/db mice, groups of 6-8 male db/db and normal db/m mice were intraperitoneal (i.p.) injected with solvent (DMSO) diluted SIS3 (Selleck Chemicals, Houston, TX, USA; cat# S7959) at dosages of 1.25, 2.5, 5mg/kg.bw/day for 8 weeks from the age of 4 weeks to 12 weeks.

Techniques: Immunostaining, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: International journal of biological sciences

Article Title: Treatment for type 2 diabetes and diabetic nephropathy by targeting Smad3 signaling.

doi: 10.7150/ijbs.87820

Figure Lengend Snippet: Figure 4. Treatment with SIS3 in db/db mice from the age of 8 weeks to 16 weeks shows less effective in controlling hyperglycemia with little improvement in glucose and insulin tolerance but effectively inhibits renal functional injury in db/db mice. (A) Fasting blood glucose. (B) Glucose tolerance test (IPGTT). (C) Insulin tolerance (IPITT). (D) HbA1c levels at 16 weeks. (E) Serum creatinine at 16 weeks. (F) Urinary albumin/creatinine ratio (UACR) at 16 weeks. Data represents the mean ± SEM for at least 6 mice per group. **P<0.001, ***P<0.001, ****P<0.0001 versus normal db/m mice. #P<0.05, ##P<0.01 compared with the control-treated db/db mice (vehicle). &&&&P<0.0001: untreated db/db mice versus normal db/m mice. $$$$P<0.0001: vehicle-treated db/db mice versus normal db/m mice. @@@@P<0.0001: SIS3-treated db/db mice versus normal db/m mice. SIS3 dosages used: SIS3 =SIS3 2.5 mg/kg.bw.

Article Snippet: To determine a safe and effective dose of SIS3 for treatment of db/db mice, groups of 6-8 male db/db and normal db/m mice were intraperitoneal (i.p.) injected with solvent (DMSO) diluted SIS3 (Selleck Chemicals, Houston, TX, USA; cat# S7959) at dosages of 1.25, 2.5, 5mg/kg.bw/day for 8 weeks from the age of 4 weeks to 12 weeks.

Techniques: Functional Assay, Control

Journal: International journal of biological sciences

Article Title: Treatment for type 2 diabetes and diabetic nephropathy by targeting Smad3 signaling.

doi: 10.7150/ijbs.87820

Figure Lengend Snippet: Figure 5. Treatment with SIS3 in db/db mice from the age of 8 weeks to 16 weeks significantly inhibits renal fibrosis in db/db mice. (A) PAS and immunostaining with antibodies against collagens I and IV, fibronectin, and α-SMA in the kidney of db/m and db/db mice treated with or without SIS3 at a dose of 2.5 mg/kg/day. (B-F) Semi-quantitation of glomerular matrix deposition and accumulation of collagens I, IV, fibronectin, and α-SMA+ myofibroblasts from the immunostained sections. (G-J) Western blotting and quantitative analysis of renal fibrosis including collagen I, α-SMA, and fibronectin. (K-M) Real-time PCR analysis for mRNA expression of fibronectin, collagen I, and α-SMA in the kidney of db/m or db/db mice at the age of 16 weeks. Data represents the mean ± SEM for at least 6 mice per group. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 versus normal db/m mice. #P<0.05, ##P<0.01, ####P<0.0001 compared with the control-treated db/db mice (vehicle). SIS3 dosages used: SIS3=SIS3 2.5 mg/kg.bw. Scale bar, 50μm.

Article Snippet: To determine a safe and effective dose of SIS3 for treatment of db/db mice, groups of 6-8 male db/db and normal db/m mice were intraperitoneal (i.p.) injected with solvent (DMSO) diluted SIS3 (Selleck Chemicals, Houston, TX, USA; cat# S7959) at dosages of 1.25, 2.5, 5mg/kg.bw/day for 8 weeks from the age of 4 weeks to 12 weeks.

Techniques: Immunostaining, Quantitation Assay, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Control

Journal: International journal of biological sciences

Article Title: Treatment for type 2 diabetes and diabetic nephropathy by targeting Smad3 signaling.

doi: 10.7150/ijbs.87820

Figure Lengend Snippet: Figure 6. Treatment of db/db mice with SIS3 from the age of 8 weeks to 16 weeks significantly inhibits renal inflammation in db/db mice. (A-C) Immunostaining and quantitative analysis of F4/80+ macrophages and expression of MCP-1 in the kidney of db/m and db/db mice at the age of 16 weeks of age. (D-F) Real-time PCR analysis detects IL-1β, TNF-α, and MCP-1 mRNA expression in the kidney of db/m and db/db mice at 16 weeks. (G-I) ELISA analysis detects serum levels of IL-1β, TNF-α and MCP-1 in db/m and db/db mice at 16 weeks. Data represents the mean ± SEM for at least 6 mice per group. *P<0.05, **P<0.01, ****P<0.0001 versus normal db/m mice. #P<0.05,

Article Snippet: To determine a safe and effective dose of SIS3 for treatment of db/db mice, groups of 6-8 male db/db and normal db/m mice were intraperitoneal (i.p.) injected with solvent (DMSO) diluted SIS3 (Selleck Chemicals, Houston, TX, USA; cat# S7959) at dosages of 1.25, 2.5, 5mg/kg.bw/day for 8 weeks from the age of 4 weeks to 12 weeks.

Techniques: Immunostaining, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: International journal of biological sciences

Article Title: Treatment for type 2 diabetes and diabetic nephropathy by targeting Smad3 signaling.

doi: 10.7150/ijbs.87820

Figure Lengend Snippet: Figure 7. Prediabetic treatment of db/db mice with SIS3 from the age of 4 weeks to 12 weeks restores the balance of Smad3/7 signaling, inactivates NF-kB/p65, and inhibits lncRNAs Erbb4-IR and LRNA9884 in the diabetic kidney of db/db mice. (A-D) Immunostaining and semi-quantitative analysis for phospho-Smad3 (p-Smad3), Smad7, and p-p65 NF-kB subunit in the kidney of db/m and db/db mice at 12 weeks. (E-J) Western blots and quantitative analysis for p-Smad3, Smurf2, Smad7, p-p65, and p-IkBα in the kidney of db/m and db/db mice at 12 weeks. (K-M) Real-time PCR analysis detects Smad7 mRNA, lncRNA Erbb4-IR, and LRNA9884 expression in the kidney of db/m and db/db mice at 12 weeks. Data represents the mean ± SEM for at least 6 mice per group. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 versus normal db/m mice. #P<0.05, ##P<0.01, ###P<0.001, ####P<0.0001 compared with the control-treated db/db mice (vehicle). SIS3 dosages used: SIS3 =SIS3 2.5 mg/kg.bw. Scale bar, 50μm.

Article Snippet: To determine a safe and effective dose of SIS3 for treatment of db/db mice, groups of 6-8 male db/db and normal db/m mice were intraperitoneal (i.p.) injected with solvent (DMSO) diluted SIS3 (Selleck Chemicals, Houston, TX, USA; cat# S7959) at dosages of 1.25, 2.5, 5mg/kg.bw/day for 8 weeks from the age of 4 weeks to 12 weeks.

Techniques: Immunostaining, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Control

Journal: International journal of biological sciences

Article Title: Treatment for type 2 diabetes and diabetic nephropathy by targeting Smad3 signaling.

doi: 10.7150/ijbs.87820

Figure Lengend Snippet: Figure 8. Treatment of db/db mice with SIS3 from the age of 8 weeks to 16 weeks restores the balance of Smad3/7 signaling, inactivates NF-kB/p65, and inhibits lncRNAs Erbb4-IR and LRNA9884 in the diabetic kidney of db/db mice. (A-D) Immunostaining and semi-quantitative analysis for phospho-Smad3 (p-Smad3), Smad7, and p-p65 NF-kB subunit in the kidney of db/m and db/db mice at 16 weeks. (E-J) Western blots and quantitative analysis for Smurf2, Smad7, p-Smad3, p-p65, and p-IkBα in the kidney of db/m and db/db mice at 16 weeks. (K-M) Real-time PCR analysis detects Smad7 mRNA, lncRNA Erbb4-IR, and LRNA9884 expression in the kidney of db/m and db/db mice at 16 weeks. Data represents the mean ± SEM for at least 6 mice per group. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 versus normal db/m mice. #P<0.05,

Article Snippet: To determine a safe and effective dose of SIS3 for treatment of db/db mice, groups of 6-8 male db/db and normal db/m mice were intraperitoneal (i.p.) injected with solvent (DMSO) diluted SIS3 (Selleck Chemicals, Houston, TX, USA; cat# S7959) at dosages of 1.25, 2.5, 5mg/kg.bw/day for 8 weeks from the age of 4 weeks to 12 weeks.

Techniques: Immunostaining, Western Blot, Real-time Polymerase Chain Reaction, Expressing

Journal: International journal of biological sciences

Article Title: Treatment for type 2 diabetes and diabetic nephropathy by targeting Smad3 signaling.

doi: 10.7150/ijbs.87820

Figure Lengend Snippet: Figure 9. Differential effect of prediabetic versus late diabetic treatment with SIS3 on islet Pax6 expression and insulin-producing b cell function in the pancreas islets of db/db mice. (A) Immunohistochemistry and multiplex immunofluorescence for detecting p-Smad3, Pax6 and insulin-producing b cells in the pancreas islets of db/m and db/db mice that receive SIS3 (2.5 mg/kg/day) treatment from the prediabetic stage at the age of 4 weeks to 12 weeks. (B) Immunohistochemistry and multiplex immunofluorescence for detecting p-Smad3, Pax6 and insulin-producing b cells in the pancreas islets of db/m and db/db mice that receive SIS3 (2.5 mg/kg/day) treatment from the age of 8 weeks to 16 weeks. (C) Quantitative analysis of p-Smad3, insulin-producing islet cells, and Pax6+ islet cells from the prediabetic-treated db/db mice. (D) Quantitative analysis of p-Smad3, insulin-producing islet cells, and Pax6+ islet cells from the late SIS3-treated db/db mice. Data represents the mean ± SEM for at least 6 mice per group. ***P<0.001, ****P<0.0001 versus normal db/m mice. #P<0.05, ##P<0.01, ####P<0.0001 compared with the control-treated db/db mice (vehicle). SIS3 dosages used: SIS3 =SIS3 2.5 mg/kg.bw. Scale bar, 50μm.

Article Snippet: To determine a safe and effective dose of SIS3 for treatment of db/db mice, groups of 6-8 male db/db and normal db/m mice were intraperitoneal (i.p.) injected with solvent (DMSO) diluted SIS3 (Selleck Chemicals, Houston, TX, USA; cat# S7959) at dosages of 1.25, 2.5, 5mg/kg.bw/day for 8 weeks from the age of 4 weeks to 12 weeks.

Techniques: Expressing, Cell Function Assay, Immunohistochemistry, Multiplex Assay, Immunofluorescence, Control

Journal: medRxiv

Article Title: Absence of phosphorylated CDK4, a biomarker of intrinsic resistance to CDK4/6 inhibitors in head and neck squamous cell carcinoma, can be predicted by their gene expression profile

doi: 10.1101/2025.11.10.25339594

Figure Lengend Snippet: HNSCC cell line model characterization. A. The effects of increasing concentrations of ribociclib (LEE011, white diamonds), abemaciclib (LY2835219, grey triangles) or palbociclib (PD0332991, black circles) on the S-phase entry rate in the indicated cell line (Cal27, Detroit 562 (noted Detroit), FaDu, SCC9, SCC15, SCC25 and SCC47) was measured by counting the proportion of cell nuclei labeled by a BrdU pulse. Each drug concentration was tested in triplicate wells of a 96-well plate. The proportions of BrdU-labeled nuclei in a well were estimated in 10 fields per well and averaged. For each cell line and condition, the mean proportion of BrdU-labeled nuclei in these fields were averaged between the triplicate wells and displayed as means ± SD. The values presented are from a typical experiment done at the same date with the three drugs for each cell line. The number of experiment done with independent cultures of each cell line are reported in the Dataset EV3. The corresponding cell line CDK4 modification profile determined by 2D gel electrophoresis of total protein extracts revealed by Western blotting with an anti-CDK4 antibody are displayed as insert in the dose response graphs or below the images (SCC90). The phosphorylated CDK4 form is indicated by an arrow. As the UM-SCC090 cells (noted SCC90) grow as 3D clumps, staining image focalization was insufficient for the BrdU labeling quantification using the ImageJ script developed to this end. Instead, representative staining images are displayed. BrdU-labeled nuclei are indicated by the white arrows. The scale bars in the top left corners of the images indicates 100 µm. The UM-SCC090 CDK4 modification profile determined by 2D gel electrophoresis of total protein extract revealed by Western blot with an anti-CDK4 antibody is displayed below the images. A single protein extract was analyzed with a single 2D gel electrophoresis for each cell line. B.The impact of increasing concentrations of the indicated drugs on the mitochondrial activity measured by the MTT cell viability assay in the 8 tested cell lines are illustrated. Drugs codes are the same as in panel A. Data are means ± SD of the MTT activity relative to the control conditions computed with data from 3 independent experiments.

Article Snippet: Palbociclib (PD0332991; S1116), abemaciclib (LY2835219; S7158) and ribociclib (LEE011; S7440) were purchased from SelleckChem (Houston, TX, USA) and dissolved in DMSO.

Techniques: Labeling, Concentration Assay, Modification, Two-Dimensional Gel Electrophoresis, Electrophoresis, Western Blot, Staining, Activity Assay, Viability Assay, Control