Journal: Bioactive Materials

Article Title: Dynamic hydrogels orchestrate the differentiation fate of mesenchymal stem cells through epigenetic regulation of SETD7 to accelerate bone defect repair

doi: 10.1016/j.bioactmat.2026.01.019

Figure Lengend Snippet: SETD7-mediated β-catenin methylation enhances its nuclear accumulation and requires binding to nuclear YAP for osteogenic activity. (a) GO analysis of predicted miRNA targets (top 20 terms). (b) Co-IP of cytoplasmic β-catenin complexes from hMSCs in HA-CA and HA-ADA hydrogels, immunoblotted for SETD7 and methyl-lysine. (c) Nuclear β-catenin and H3 levels. (d, e) Quantification of (b) and (c). (f) Co-IP analysis following SETD7 overexpression (MOI = 20, 12 h transduction, 48 h culture). (g) Nuclear β-catenin levels after SETD7 overexpression. (h, i) Quantification of (f) and (g). (j) Co-IP analysis following SETD7 knockdown (MOI = 10, 24 h transduction, 48 h culture). (k) Nuclear β-catenin levels after SETD7 silencing. (l, m) Quantification of (j) and (k). (n, o) Nuclear YAP and H3 expression with quantification. (p, q) Co-IP analysis of nuclear YAP-β-catenin interaction in hMSCs cultured in HA-CA and HA-ADA hydrogels. (r) β-catenin/YAP co-localization by immunofluorescence. (s) β-catenin and YAP staining after gene silencing. (t, u) ARS staining and quantification of osteogenic differentiation following knockdown. n = 3. Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: After blocking with 5 % non-fat milk for 1 h at room temperature, membranes were incubated with primary antibodies including β-catenin (1:800, Santa Cruz Biotechnology, #sc-7963), YAP (1:1000, ABclonal, #A19134), SETD7 (1:1000, Proteintech, #24840-1-AP), methylated lysine (1:1200, Abcam, #ab23366), Runx2 (1:1000, ABclonal, #A2851), OCN (1:1000, ABclonal, #A20800), and PPARγ (1:1000, ABclonal, #A11183) overnight at 4 °C, followed by HRP-conjugated secondary antibodies for 1 h at room temperature.

Techniques: Methylation, Binding Assay, Activity Assay, Co-Immunoprecipitation Assay, Over Expression, Transduction, Knockdown, Expressing, Cell Culture, Immunofluorescence, Staining

Journal: Bioactive Materials

Article Title: Dynamic hydrogels orchestrate the differentiation fate of mesenchymal stem cells through epigenetic regulation of SETD7 to accelerate bone defect repair

doi: 10.1016/j.bioactmat.2026.01.019

Figure Lengend Snippet: Schematic illustration of the HA-ADA hydrogel system for enhanced bone regeneration through dynamic matrix-mediated mechanotransduction. (a) Experimental workflow. (b) Chemical composition of hydrogels showing HA-ADA or HA-CA polymers decorated with RGD motifs, which crosslink with Ac-β-CD under blue light to form distinct network architectures. (c) Comparative illustration of hydrogel performance in bone regeneration. HA-ADA hydrogels exhibit high network adaptability that enables cell spreading through dynamic “gate opening” mechanisms involving actin polymerization and F-actin bundle formation, leading to enhanced bone regeneration. In contrast, HA-CA hydrogels display low network adaptability with restricted “gate closed” states that limit cell spreading and result in reduced bone regeneration capacity. (d) Molecular mechanism underlying HA-ADA hydrogel-promoted osteogenesis. The dynamic matrix downregulates miR-376a-3p and miR-127-5p expression, thereby relieving their repression of SETD7. Elevated SETD7 subsequently methylates β-catenin, facilitating YAP/β-catenin signaling cascade activation to drive osteogenic differentiation.

Article Snippet: After blocking with 5 % non-fat milk for 1 h at room temperature, membranes were incubated with primary antibodies including β-catenin (1:800, Santa Cruz Biotechnology, #sc-7963), YAP (1:1000, ABclonal, #A19134), SETD7 (1:1000, Proteintech, #24840-1-AP), methylated lysine (1:1200, Abcam, #ab23366), Runx2 (1:1000, ABclonal, #A2851), OCN (1:1000, ABclonal, #A20800), and PPARγ (1:1000, ABclonal, #A11183) overnight at 4 °C, followed by HRP-conjugated secondary antibodies for 1 h at room temperature.

Techniques: Expressing, Activation Assay

Journal: Non-coding RNA Research

Article Title: Exosomal miRNA-218–5p derived from low-passage dermal papilla cells modulates hair follicle growth and development

doi: 10.1016/j.ncrna.2026.01.004

Figure Lengend Snippet: MiR-218-5p regulated HF growth- and development-related gene expression in HFSCs. (A) MiR-218–5p expression levels in HFSCs after transfection with miR-218–5p mimics or the inhibitor (unpaired two-tailed t -test, n = 3). (B) Expression of HF development-related genes in HFSCs is regulated by miR-218–5p. (C) β-Catenin and SFRP2 protein expression in HFSCs after treatment with miR-218–5p mimics or inhibitor (unpaired two-tailed t -test, n = 3). ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: Anti-SFRP2 rabbit polyclonal antibody (Proteintech Biotech, Cat No. 12189-1-AP), anti-β-catenin polyclonal antibody (Proteintech, China, Cat No. 51067-2-AP), and anti-GAPDH mouse monoclonal antibody (Proteintech, China, Cat No. 60004-1-Ig) were the primary antibodies.

Techniques: Gene Expression, Expressing, Transfection, Two Tailed Test

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 increases β-Catenin activity and promotes its nuclear translocation (A) Western blot shows Panx1 overexpression and knockout efficiency (GAPDH as loading control). (B) Glycosylation patterns of exogenous Panx1. (C and D) Total and non-phosphorylated β-catenin levels in whole cell and nuclear fractions (Lamin B1 as loading control). (E) Immunofluorescence of β-catenin nuclear translocation (red; nuclei, blue). Scale bars, 25 μm. (F) qRT-PCR of Wnt/β-catenin downstream gene expression normalized to PPIA. (G) Western blot of Wnt/β-catenin downstream proteins (GAPDH as loading control). Data are mean ± SD, statistics in (C), (D), (F), and (G): ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, two-sided Student’s t test.

Article Snippet: β-Catenin (D10A8) XP Rabbit mAb , Cell Signaling Technology , Cat# 8480S; RRID: AB_11127855.

Techniques: Activity Assay, Translocation Assay, Western Blot, Over Expression, Knock-Out, Control, Glycoproteomics, Immunofluorescence, Quantitative RT-PCR, Gene Expression

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 forms complexes with β-Catenin and GSK-3β (A) Western blot of destruction complex proteins in Panx1-OE and Panx1 −/− 3T3-L1 cells (GAPDH as loading control). (B) Co-IP of Panx1 with β-catenin or destruction complex proteins in HEK293T cells expressing Panx1-HA. (C) Co-IP of β-catenin (total or phosphorylated) with GSK-3β in Panx1-OE and Panx1 −/− cells (GAPDH as loading control for input). (D) Western blot showing β-catenin knockdown efficiency in Panx1-OE cells (GAPDH as loading control). (E) Co-IP of Panx1 with GSK-3β after β-catenin knockdown (GAPDH as loading control for input). (F) Predicted 3D structures of Panx1, β-catenin, and GSK-3β complexes showing conformational changes. (G) Ranking scores of binding affinities among Panx1, β-catenin, and GSK-3β. Data are mean ± SD, statistics in (A): ∗ p < 0.05 and ∗∗ p < 0.01, two-sided Student’s t test.

Article Snippet: β-Catenin (D10A8) XP Rabbit mAb , Cell Signaling Technology , Cat# 8480S; RRID: AB_11127855.

Techniques: Western Blot, Control, Co-Immunoprecipitation Assay, Expressing, Knockdown, Binding Assay

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Effect of Panx1 on cell proliferation (A) Cell viability of Panx1-OE and Panx1 −/− cells. (B–D) Flow cytometry analysis of cell cycle distribution (PI staining) with the quantification of G1, S, and G2/M phases. (E) Western blot confirms β-catenin knockdown and effects on downstream proteins in Panx1-OE cells. (F and G) Flow cytometry and quantification of cell cycle distribution after β-catenin knockdown. Each panel represents an independently generated pair of stable lines, and statistical analyses are performed within each matched pair. Data are mean ± SD, statistics in (A), (D), and (G): ∗∗ p < 0.01 and ∗∗∗ p < 0.001, two-sided Student’s t test.

Article Snippet: β-Catenin (D10A8) XP Rabbit mAb , Cell Signaling Technology , Cat# 8480S; RRID: AB_11127855.

Techniques: Flow Cytometry, Staining, Western Blot, Knockdown, Generated

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 knockout downregulated the Wnt/β-Catenin pathway in eWAT tissue (A) PCA of gene expression profiles ( n = 6/group). (B) GSEA enrichment of the Wnt/β-catenin pathway in eWAT. (C) Venn diagram showing overlap of DEGs with Wnt/β-catenin pathway genes. (D) Heatmap of Wnt-related DEGs in WT and Panx1 −/− mice. (E) Western blot of active β-catenin and downstream proteins (β-actin as loading control). Data are mean ± SD, statistics in (E): ∗ p < 0.05, ∗∗ p < 0.01, two-sided Student’s t test.

Article Snippet: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) Rabbit mAb , Cell Signaling Technology , Cat# 8814S; RRID: AB_11127856.

Techniques: Knock-Out, Gene Expression, Western Blot, Control

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 increases β-Catenin activity and promotes its nuclear translocation (A) Western blot shows Panx1 overexpression and knockout efficiency (GAPDH as loading control). (B) Glycosylation patterns of exogenous Panx1. (C and D) Total and non-phosphorylated β-catenin levels in whole cell and nuclear fractions (Lamin B1 as loading control). (E) Immunofluorescence of β-catenin nuclear translocation (red; nuclei, blue). Scale bars, 25 μm. (F) qRT-PCR of Wnt/β-catenin downstream gene expression normalized to PPIA. (G) Western blot of Wnt/β-catenin downstream proteins (GAPDH as loading control). Data are mean ± SD, statistics in (C), (D), (F), and (G): ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, two-sided Student’s t test.

Article Snippet: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) Rabbit mAb , Cell Signaling Technology , Cat# 8814S; RRID: AB_11127856.

Techniques: Activity Assay, Translocation Assay, Western Blot, Over Expression, Knock-Out, Control, Glycoproteomics, Immunofluorescence, Quantitative RT-PCR, Gene Expression

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Panx1 forms complexes with β-Catenin and GSK-3β (A) Western blot of destruction complex proteins in Panx1-OE and Panx1 −/− 3T3-L1 cells (GAPDH as loading control). (B) Co-IP of Panx1 with β-catenin or destruction complex proteins in HEK293T cells expressing Panx1-HA. (C) Co-IP of β-catenin (total or phosphorylated) with GSK-3β in Panx1-OE and Panx1 −/− cells (GAPDH as loading control for input). (D) Western blot showing β-catenin knockdown efficiency in Panx1-OE cells (GAPDH as loading control). (E) Co-IP of Panx1 with GSK-3β after β-catenin knockdown (GAPDH as loading control for input). (F) Predicted 3D structures of Panx1, β-catenin, and GSK-3β complexes showing conformational changes. (G) Ranking scores of binding affinities among Panx1, β-catenin, and GSK-3β. Data are mean ± SD, statistics in (A): ∗ p < 0.05 and ∗∗ p < 0.01, two-sided Student’s t test.

Article Snippet: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) Rabbit mAb , Cell Signaling Technology , Cat# 8814S; RRID: AB_11127856.

Techniques: Western Blot, Control, Co-Immunoprecipitation Assay, Expressing, Knockdown, Binding Assay

Journal: iScience

Article Title: Panx1 deficiency exacerbates GAN diet-induced obesity by destabilizing β-catenin via GSK3β

doi: 10.1016/j.isci.2026.115098

Figure Lengend Snippet: Effect of Panx1 on cell proliferation (A) Cell viability of Panx1-OE and Panx1 −/− cells. (B–D) Flow cytometry analysis of cell cycle distribution (PI staining) with the quantification of G1, S, and G2/M phases. (E) Western blot confirms β-catenin knockdown and effects on downstream proteins in Panx1-OE cells. (F and G) Flow cytometry and quantification of cell cycle distribution after β-catenin knockdown. Each panel represents an independently generated pair of stable lines, and statistical analyses are performed within each matched pair. Data are mean ± SD, statistics in (A), (D), and (G): ∗∗ p < 0.01 and ∗∗∗ p < 0.001, two-sided Student’s t test.

Article Snippet: Non-phospho (Active) β-Catenin (Ser33/37/Thr41) Rabbit mAb , Cell Signaling Technology , Cat# 8814S; RRID: AB_11127856.

Techniques: Flow Cytometry, Staining, Western Blot, Knockdown, Generated