Journal: Bone Research

Article Title: KDM6B safeguards mineralized tissue homeostasis from mechanical stress through epigenetic control of PIEZO1-mediated mechanotransduction in the mouse incisor

doi: 10.1038/s41413-026-00544-2

Figure Lengend Snippet: KDM6B epigenetically regulates BMI1 to silence PIEZO1 expression and maintain tissue homeostasis. a CUT&RUN profiles of H3K27me3 in the proximal incisor region of control and Gli1-CreER T2 ;Kdm6b fl/fl mice at 5 dpt. TSS: Transcription start sites. TES: Transcription end sites. b Venn diagram showing overlap between genes with increased H3K27me3 enrichment in CUT&RUN and downregulated genes in RNA-seq from Gli1-CreER T2 ;Kdm6b fl/fl incisors. c Heatmap of overlapping genes expressed in TACs in control and Gli1-CreER T2 ;Kdm6b fl/fl mice. d Co-staining of Bmi1 (white) and Kdm6b (red) in the wild-type mouse incisors. d’ Magnified view of boxed region. White dotted lines outline the cervical loop. Yellow arrows indicate double-positive cells. Scale bars, 50 μm. e CUT&RUN tracks showing H3K27me3 enrichment at the Bmi1 locus in control and Gli1-CreER T2 ;Kdm6b fl/fl incisors. IgG serves as a negative control. Red lines indicate CRISPRi gRNA target sites. f RT-qPCR analysis of Bmi1 expression following CRISPRi targeting (mean ± SEM, n = 3; gRNA1, P = 0.034 6; gRNA2, P = 0.709 1; gRNA3, P = 0.058 7). g –i Bmi1 in situ hybridization in control, Kdm6b mutant, and Ezh2 rescue incisors. g’ –i’ Magnified views of boxed regions. White dotted lines outline the cervical loop. Yellow arrows indicate positive signals. Asterisk indicates absence of signal. j Relative Bmi1 mRNA expression in the three genotypes (mean ± SEM, n = 3; control vs Kdm6b mutant, P = 0.004 2; Kdm6b mutant vs Ezh2 rescue, P = 0.000 2; control vs Ezh2 rescue, P = 0.012 8). k Piezo1 expression in dental mesenchymal cells after 3 days of control or Bmi1 siRNA treatment (mean ± SEM, n = 3; P = 0.036 4). l Piezo1 expression after vector or Bmi1 plasmid treatment (mean ± SEM, n = 3; ctrl+vector vs. ctrl+ Bmi1 , P = 0.022 6; ctrl+vector vs. mutant+vector, P = 0.000 8; ctrl+vector vs. mutant+ Bmi1 , P = 0.838 4; mutant+vector vs. mutant+ Bmi1 , P = 0.000 9). m CUT&RUN profiles showing BMI1 enrichment at the Piezo1 locus (IgG negative control and two BMI1 replicates). Red lines indicate CRISPRi gRNA target sites. n RT-qPCR analysis of Piezo1 expression following CRISPRi targeting of BMI1-binding regions (mean ± SEM, n = 3; gRNA1, P = 0.012 3 ; gRNA2, P = 0.029 7). Kdm6b mutant: Gli1-CreER T2 ;Kdm6b fl/fl . Ezh2 rescue: Gli1-CreER T2 ;Kdm6b fl/fl ;Ezh2 fl/+

Article Snippet: For plasmid transfection, cells were transfected with 1 μg/μL Bmi1 (Myc-DDK-tagged) plasmid (OriGene, MR226936; vector, PS100001 ) DNA using LipofectamineTM 3000 Transfection Reagent (Thermo Fisher Scientific, L3000150) for 3 days, followed by TUNEL staining and qPCR.

Techniques: Expressing, Control, RNA Sequencing, Staining, Negative Control, Quantitative RT-PCR, In Situ Hybridization, Mutagenesis, Plasmid Preparation, Binding Assay

Journal: Bone Research

Article Title: KDM6B safeguards mineralized tissue homeostasis from mechanical stress through epigenetic control of PIEZO1-mediated mechanotransduction in the mouse incisor

doi: 10.1038/s41413-026-00544-2

Figure Lengend Snippet: Schematic representation of KDM6B safeguarding tissue homeostasis to mechanical stress through epigenetic control of PIEZO1-mediated mechanotransduction. Using the mouse incisor as a model of mechanical loading, we reveal that within TACs, Kdm6b demethylates H3K27me3, thereby relieving the repression of the Bmi1 gene. Normal BMI1 inhibits Piezo1 expression. This maintains physiological PIEZO1 levels, ensuring calibrated Ca 2+ influx for proliferation and differentiation. In contrast, loss of Kdm6b leads to an accumulation of H3K27me3 at the Bmi1 promoter region, which silences Bmi1 expression and diminishes BMI1 formation. This reduction results in pathologically increased PIEZO1 ion channels in the membrane. The subsequent Ca 2+ overload triggers TAC apoptosis while reducing proliferation and differentiation. Ultimately, these molecular events compromise tissue homeostasis. Schematic created with BioRender.com. Ho, T. (2026) https://BioRender.com/8mzv4a3

Article Snippet: For plasmid transfection, cells were transfected with 1 μg/μL Bmi1 (Myc-DDK-tagged) plasmid (OriGene, MR226936; vector, PS100001 ) DNA using LipofectamineTM 3000 Transfection Reagent (Thermo Fisher Scientific, L3000150) for 3 days, followed by TUNEL staining and qPCR.

Techniques: Control, Expressing, Membrane

Journal: Bone Research

Article Title: KDM6B safeguards mineralized tissue homeostasis from mechanical stress through epigenetic control of PIEZO1-mediated mechanotransduction in the mouse incisor

doi: 10.1038/s41413-026-00544-2

Figure Lengend Snippet: KDM6B epigenetically regulates BMI1 to silence PIEZO1 expression and maintain tissue homeostasis. a CUT&RUN profiles of H3K27me3 in the proximal incisor region of control and Gli1-CreER T2 ;Kdm6b fl/fl mice at 5 dpt. TSS: Transcription start sites. TES: Transcription end sites. b Venn diagram showing overlap between genes with increased H3K27me3 enrichment in CUT&RUN and downregulated genes in RNA-seq from Gli1-CreER T2 ;Kdm6b fl/fl incisors. c Heatmap of overlapping genes expressed in TACs in control and Gli1-CreER T2 ;Kdm6b fl/fl mice. d Co-staining of Bmi1 (white) and Kdm6b (red) in the wild-type mouse incisors. d’ Magnified view of boxed region. White dotted lines outline the cervical loop. Yellow arrows indicate double-positive cells. Scale bars, 50 μm. e CUT&RUN tracks showing H3K27me3 enrichment at the Bmi1 locus in control and Gli1-CreER T2 ;Kdm6b fl/fl incisors. IgG serves as a negative control. Red lines indicate CRISPRi gRNA target sites. f RT-qPCR analysis of Bmi1 expression following CRISPRi targeting (mean ± SEM, n = 3; gRNA1, P = 0.034 6; gRNA2, P = 0.709 1; gRNA3, P = 0.058 7). g –i Bmi1 in situ hybridization in control, Kdm6b mutant, and Ezh2 rescue incisors. g’ –i’ Magnified views of boxed regions. White dotted lines outline the cervical loop. Yellow arrows indicate positive signals. Asterisk indicates absence of signal. j Relative Bmi1 mRNA expression in the three genotypes (mean ± SEM, n = 3; control vs Kdm6b mutant, P = 0.004 2; Kdm6b mutant vs Ezh2 rescue, P = 0.000 2; control vs Ezh2 rescue, P = 0.012 8). k Piezo1 expression in dental mesenchymal cells after 3 days of control or Bmi1 siRNA treatment (mean ± SEM, n = 3; P = 0.036 4). l Piezo1 expression after vector or Bmi1 plasmid treatment (mean ± SEM, n = 3; ctrl+vector vs. ctrl+ Bmi1 , P = 0.022 6; ctrl+vector vs. mutant+vector, P = 0.000 8; ctrl+vector vs. mutant+ Bmi1 , P = 0.838 4; mutant+vector vs. mutant+ Bmi1 , P = 0.000 9). m CUT&RUN profiles showing BMI1 enrichment at the Piezo1 locus (IgG negative control and two BMI1 replicates). Red lines indicate CRISPRi gRNA target sites. n RT-qPCR analysis of Piezo1 expression following CRISPRi targeting of BMI1-binding regions (mean ± SEM, n = 3; gRNA1, P = 0.012 3 ; gRNA2, P = 0.029 7). Kdm6b mutant: Gli1-CreER T2 ;Kdm6b fl/fl . Ezh2 rescue: Gli1-CreER T2 ;Kdm6b fl/fl ;Ezh2 fl/+

Article Snippet: These gRNAs were cloned into the pCas-Guide-Puro-CRISPRi vector to generate a custom Bmi1 CRISPRi kit (Origene Technologies, CW312379 ), which was constructed by Origene Technologies.

Techniques: Expressing, Control, RNA Sequencing, Staining, Negative Control, Quantitative RT-PCR, In Situ Hybridization, Mutagenesis, Plasmid Preparation, Binding Assay

Journal: Bone Research

Article Title: KDM6B safeguards mineralized tissue homeostasis from mechanical stress through epigenetic control of PIEZO1-mediated mechanotransduction in the mouse incisor

doi: 10.1038/s41413-026-00544-2

Figure Lengend Snippet: Schematic representation of KDM6B safeguarding tissue homeostasis to mechanical stress through epigenetic control of PIEZO1-mediated mechanotransduction. Using the mouse incisor as a model of mechanical loading, we reveal that within TACs, Kdm6b demethylates H3K27me3, thereby relieving the repression of the Bmi1 gene. Normal BMI1 inhibits Piezo1 expression. This maintains physiological PIEZO1 levels, ensuring calibrated Ca 2+ influx for proliferation and differentiation. In contrast, loss of Kdm6b leads to an accumulation of H3K27me3 at the Bmi1 promoter region, which silences Bmi1 expression and diminishes BMI1 formation. This reduction results in pathologically increased PIEZO1 ion channels in the membrane. The subsequent Ca 2+ overload triggers TAC apoptosis while reducing proliferation and differentiation. Ultimately, these molecular events compromise tissue homeostasis. Schematic created with BioRender.com. Ho, T. (2026) https://BioRender.com/8mzv4a3

Article Snippet: These gRNAs were cloned into the pCas-Guide-Puro-CRISPRi vector to generate a custom Bmi1 CRISPRi kit (Origene Technologies, CW312379 ), which was constructed by Origene Technologies.

Techniques: Control, Expressing, Membrane

Journal: Scientific Reports

Article Title: A CXCR4 targeting peptide delivered by silica nanoparticles eliminates migrating cancer stem cells in pancreatic ductal adenocarcinoma

doi: 10.1038/s41598-026-48584-2

Figure Lengend Snippet: BMI1 downstream CXCL12–CXCR4 regulates EMT and stemness. ( A ) Protein–protein interactions of CXCL12 and CXCR4 with relevant factors involved in metastasis (red), stemness (purple), sonic hedgehog signaling (blue), AKT signaling (grey) and NFκB pathway (yellow) (STRING). ( B ) BMI1 gene expression analysis and western blot analysis. GAPDH was used as a loading control. Intensity ratios (IR) calculated against control lane using ImageJ. Cropped blot for clarity. ( C ) Gene expression analysis with genes involved in EMT using qRT-PCR. ( D ) Immunofluorescence quantifications and representative micrographs for indicated cell lines with white arrowheads marking mesenchymal structures of actin filaments stained with Phalloidin (pink) and nucleus stained with DAPI (blue). ( E ) Migration assays towards serum containing media. ( F ) Experimental scheme to evaluate CD133 and CXCR4 surface expression using flow cytometry for Panc354 and MetPO1 (sh_ SCR , sh1_ BMI1 and sh2_ BMI1 ) with (or without) CXCL12. ( G ) Flow cytometry analysis of CD133 + cells and CD133 + CXCR4 + cells in MetPO1 (sh_ SCR , sh1_ BMI1 and sh2_ BMI1 ) treated with (or without) CXCL12. ( H ) Representative cytometry plots for MetPO1 cell line. Error bars represent the standard deviation. n = 3 for all experiments unless otherwise depicted in the datasets. * p < 0.05, ns = not significant.

Article Snippet: Recombinant BMI1 IgG1 (REA438) , APC , Miltenyi Biotec , 130-124-301.

Techniques: Protein-Protein interactions, Gene Expression, Western Blot, Control, Quantitative RT-PCR, Immunofluorescence, Staining, Migration, Expressing, Flow Cytometry, Cytometry, Standard Deviation

Journal: Scientific Reports

Article Title: A CXCR4 targeting peptide delivered by silica nanoparticles eliminates migrating cancer stem cells in pancreatic ductal adenocarcinoma

doi: 10.1038/s41598-026-48584-2

Figure Lengend Snippet: JM#21, most potent EPIX4 derivative to target miCSCs. ( A ) Migration assays towards CXCL12 using Panc354 for EPI-X4 and its derivatives at depicted concentrations. Pre-treatment with EPI-X4, WSCO2, JM#21 and the inactive peptide was applied for 30 min. ( B ) Representative micrographs (10x, DAPI staining) of transwell migration assays in Panc354 cells for the indicated conditions and concentrations. ( C ) Migration assays towards CXCL12 for MetPO1 using JM#21 and the inactive peptide at depicted concentrations. ( D ) Quantifications of percent mesenchymal structures after 15 min and 6 h of CXCL12 treatment in MetPO1 cells. JM#21 pre-treatment was applied for 30 min and representative micrographs with white arrowheads marking mesenchymal structures of actin filaments stained with Phalloidin (pink) and nuclear staining using DAPI (blue). ( E ) Gene expression analysis for indicated cell lines with genes involved in EMT and SHH pathway. ( F ) Gene expression analysis for indicated cell lines with genes involved in stemness. ( G ) Sphere formation assays for 1 st and 2 nd generation of sphere formation. ( H ) Western blot analysis of CADHERIN-1, VIMENTIN, CADHERIN-2, NANOG and BMI1 for indicated cell lines. GAPDH was used as a loading control. Intensity ratios (IR) calculated against control lane using ImageJ. Cropped blot for clarity. ( I ) Experimental design for combination therapy analyzing relapse using JM#21, gemcitabine (labelled as G) and paclitaxel (labelled as P). Quantification of cell viability and representative pictures for clonogenic assays after treatment with JM#21 (10 μM ), gemcitabine (Gem) for indicated concentrations as depicted in experimental design in MetPO1 cell line. ( J ) Flow cytometry for CD133 in Panc354 and MetPO1cells for the indicated treatments shown as fold change. Error bars represent the standard deviation. n = 3 for all experiments unless otherwise depicted in the datasets. * p < 0.05, ns = not significant.

Article Snippet: Recombinant BMI1 IgG1 (REA438) , APC , Miltenyi Biotec , 130-124-301.

Techniques: Migration, Staining, Gene Expression, Western Blot, Control, Flow Cytometry, Standard Deviation