Journal: Journal of Oral Biology and Craniofacial Research

Article Title: Effect of dimethyl sulfoxide pretreatment on bonding of universal adhesives to pulp chamber dentin

doi: 10.1016/j.jobcr.2026.101409

Figure Lengend Snippet: SEM ( × 1000) photomicrograph of resin-dentin interface of Group 1A (Adper Single Bond 2 adhesive without DMSO Pretreatment): (a) Immediate; (b) Delayed – poor interfacial seal is evident at six months; Group 2A (Tetric N-Bond Universal Adhesive without DMSO Pretreatment): (c) Immediate; (d) Delayed - poor interfacial adaptation at six months. 3A (Prime & Bond Universal without DMSO Pretreatment): (e) Immediate; (f) Delayed - interfacial gap is evident at six months. Group 1B (Adper Single Bond 2 adhesive with DMSO Pretreatment): (g) Immediate; (h) Delayed - interfacial seal is seen at both time periods. Group 2B (Tetric N-Bond Universal Adhesive with DMSO Pretreatment): (i) Immediate; (j) Delayed - good interfacial adaptation can be seen even at six months. Group 3B (Prime & Bond Universal Adhesive with DMSO Pretreatment): (a) Immediate; (b) Delayed - good interfacial seal is present even after six months.

Article Snippet: Group 1A (ASB) Adper Single Bond 2 adhesive (3M ESPE): Acid etching with 36 % phosphoric acid (Detrey conditioner 36, Dentsply Sirona) for 15 s was done followed by rinsing with water for 10 s and then blot dried.

Techniques: Adhesive

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Soluble VCAM-1 May Serve as a Pharmacodynamic CSF Marker to Monitor BACE2 Activity in Non-Human Primates

doi: 10.1016/j.mcpro.2025.101012

Figure Lengend Snippet: Western blot validation of VCAM-1 as BACE2 substrate. A , scheme of VCAM-1 cleavage by BACE2. Mapping of detected peptides ( B ) suggests that VCAM-1 is shed closely above the plasma membrane. Western blotting was conducted with a polyclonal antibody that specifically targets the ectodomain. The figure was created with BioRender.com . B , Western blot detection of sVCAM-1 in CSF from cynomolgus monkeys treated with IgG RSV, anti-BACE1, ATV:BACE1 or verubecestat (N = 4 in each group). Albumin served as loading control and was visualized by total protein staining. C , quantification of Western blot results in B . First, sVCAM-1 signal intensities were normalized to albumin. For each individual, the two pre-dose values were averaged, followed by calculation of the post-dose/pre-dose ratio. Data depict mean and SD. One-way ANOVA in combination with Tukey’s multiple comparisons test. Only significant differences are indicated. ∗ p < 0.05, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The peptide was dissolved in 50 mM sodium acetate pH 4.4, and 0.5 μg of the peptide were incubated with 0.5 μg recombinant human BACE2 (R&D Systems) overnight at 37 °C.

Techniques: Western Blot, Biomarker Discovery, Clinical Proteomics, Membrane, Control, Staining

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Soluble VCAM-1 May Serve as a Pharmacodynamic CSF Marker to Monitor BACE2 Activity in Non-Human Primates

doi: 10.1016/j.mcpro.2025.101012

Figure Lengend Snippet: Validation of VCAM-1 as BACE2 substrate in human CSF . A , Western blot detection of sVCAM-1 in CSF from participants of a clinical phase 1 study . Participants were either treated with placebo (N = 6) or with 550 mg verubecestat (N = 5), and CSF was sampled 2 h before (pre-dose, pre) and 24 h after treatment (post-dose, post). Albumin served as loading control and was visualized by total protein staining. B , quantification of Western blot results in A . sVCAM-1 signal intensities were first normalized to albumin. Then, the post-dose value for each individual was normalized to the corresponding pre-dose value. Data show mean and SD. Unpaired Student’s t test. ∗∗∗ p < 0.001.

Article Snippet: The peptide was dissolved in 50 mM sodium acetate pH 4.4, and 0.5 μg of the peptide were incubated with 0.5 μg recombinant human BACE2 (R&D Systems) overnight at 37 °C.

Techniques: Biomarker Discovery, Western Blot, Control, Staining

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Soluble VCAM-1 May Serve as a Pharmacodynamic CSF Marker to Monitor BACE2 Activity in Non-Human Primates

doi: 10.1016/j.mcpro.2025.101012

Figure Lengend Snippet: Identification of the BACE2 cleavage site in VCAM-1 . A , localization and length of sVCAM-1 peptides detected in NHP CSF (N = 4) after GluC digest in a semi-specific TermineR-based search using the Fragpipe software. Peptides were mapped to the UniProt full-length Macaca fascicularis VCAM-1 sequence. Protein domain annotations were annotated based on prediction by Phobius. Pre-dose and post-dose CSF from the verubecestat treatment group was analyzed, and peptide LFQ values in the post-dose samples were normalized to the corresponding pre-dose sample. Peptide coloring represents the log 2 post-dose/pre-dose ratio. Coloring in dark blue or dark red indicates peptides only detected in pre-dose or post-dose CSF, respectively. Peptides either detected in pre-dose or post-dose CSF, but not in both samples of the same animal (so that calculation of the individual post-dose/pre-dose ratio was not possible), are colored in dark blue and dark red . Unspecific peptide termini, not resulting from GluC cleavage, are highlighted by yellow triangles . B , sVCAM-1 peptide mapping for the same samples as in A after analysis in Spectronaut. C , the semi-specific peptide identified in A and B , which potentially marks the BACE2 cleavage site, aligns with both Macaca fascicularis and human VCAM-1. Ectodomain ( blue ) and transmembrane domain ( yellow ) annotation is based on Phobius prediction (NHP) and UniProt (human), respectively. Amino acids in the human sequence that differ from the NHP VCAM-1 sequence are colored in red . In an in vitro cleavage assay, a synthetic VCAM-1 peptide was cleaved by BACE2 at the same position as found for CSF sVCAM-1. D , total ion chromatograms of the BACE2 cleavage assay of the synthetic VCAM-1 peptide depicted in C . Total ion chromatograms are shown for one representative out of two experiments. E , MS2 spectrum for the BACE2-cleaved synthetic VCAM-1 peptide. F , MS2 spectrum (hyperscore = 24.7) for the semi-specific, potentially BACE2-cleaved VCAM-1 peptide identified in NHP CSF. G , sequence logo graphic generated with WebLogo of the amino acid alignment of published BACE1 and 2 substrates around the published cleavage sites . Positively charged amino acids are displayed in blue , negatively charged amino acids in red , polar amino acids and glycine in green or purple (glutamine, asparagine), and non-polar amino acids are shown in black . The height of the amino acid symbols indicates their frequency at this position.

Article Snippet: The peptide was dissolved in 50 mM sodium acetate pH 4.4, and 0.5 μg of the peptide were incubated with 0.5 μg recombinant human BACE2 (R&D Systems) overnight at 37 °C.

Techniques: Software, Sequencing, In Vitro, Cleavage Assay, Generated

Journal: Cell Death & Disease

Article Title: Asb10 accelerates pathological cardiac remodeling by stabilizing HSP70

doi: 10.1038/s41419-025-07735-5

Figure Lengend Snippet: A Principal component analysis (PCA) of merged GEO datasets. N = 11. B Volcano plot of the merged GEO datasets. N = 11. C Heat map showing the relative expression of Asb10 and hypertrophic markers in merged GEO datasets. N = 11. D Validation of the relative Asb10 expression in cardiac tissue of sham and TAC mice from three GEO datasets, GSE79536 , GSE136308 and GSE180720 . N = 3 for GSE79536 and GSE180720 , N = 5 (sham) and 4 (TAC) in GSE136308 . E Validation of the relative Asb10 expression in cardiac tissue of donors, hypertrophic cardiomyopathy and heart failure patients from three GEO datasets, GES180313, GSE135055 , GSE161472 . N = 7 (Donor) and 13 (HCM) in GSE180313 , N = 9 (Donor) and 13 (HF) in GSE135055 , N = 4 (NF) and 5 (HF) in GSE161472 . F In vitro validation of the protein expression of ASB10 in NRVMs treated with 50 μM PE for 24 h as determined by western blot. The right panel showed the densitometric quantification. N = 9. G In vitro validation of the mRNA expression of Asb10 in NRVMs treated with 50 μM PE for 24 h as determined by qPCR. N = 6. H Representative images of anti-ASB10 immunofluorescence staining in NRVMs treated with 50 μM PE for 24 h. Scale bar = 10 μm. The right panel showed the quantification. N = 6. I In vivo validation of the protein expression of ASB10 in heart tissue from mice treated with TAC surgery as determined by western blot. The right panel showed the densitometric quantification. N = 5. J In vivo validation of the mRNA expression of Asb10 in heart tissues from mice treated with TAC surgery as determined by qPCR. N = 5. Limma power differential expression analysis ( B – E ) was conducted. Unpaired Student’s t -test ( F , G ) was conducted. One-way ANOVA followed by Turkey’s multiple comparisons test was conducted ( I , J ). Data were represented as Mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns P > 0.05.

Article Snippet: Antibody against Asb10 (#sc-100677) was purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Expressing, Biomarker Discovery, In Vitro, Western Blot, Immunofluorescence, Staining, In Vivo, Quantitative Proteomics

Journal: Cell Death & Disease

Article Title: Asb10 accelerates pathological cardiac remodeling by stabilizing HSP70

doi: 10.1038/s41419-025-07735-5

Figure Lengend Snippet: NRVMs were infected with adenovirus-packed Asb10 or Vector for 30 h, then treated with 50 μM PE for 24 h, and cells were then collected for further detection. A Protein expression of hypertrophic markers with GAPDH as loading control as determined by western blot. The right panel showed the densitometric quantifications. N = 6. B Relative mRNA expressions of hypertrophic markers as determined by qPCR. N = 6. C Representative images of anti-α-actinin immunofluorescence staining showing cell size. The right panel showed the quantification. Scale bar = 30 μm. N = 56 (Ad- Vector +Vehicle), 54 (Ad- Vector + PE), 50 (Ad- Asb10 +Vehicle), 53 (Ad- Asb10 + PE). Two-way ANOVA followed by Turkey’s multiple comparisons test was conducted. Data were represented as Mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Antibody against Asb10 (#sc-100677) was purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Infection, Plasmid Preparation, Expressing, Control, Western Blot, Immunofluorescence, Staining

Journal: Cell Death & Disease

Article Title: Asb10 accelerates pathological cardiac remodeling by stabilizing HSP70

doi: 10.1038/s41419-025-07735-5

Figure Lengend Snippet: A ASB10 interactome from BioGRID ( www.thebiogrid.org ). B Immunoprecipitation-mass spectrometry assay to screen the Asb10 interactome in NRVMs overexpressing Asb10. The top 20 interacting proteins were shown in the upper panel, and the peptide map of the HSPA1a peptide with the highest mapping score was shown in the lower panel. C Co-immunoprecipitation assay to validate the interacting proteins in NRVMs. N = 3. D Co-immunoprecipitation assay to validate the interaction between ASB10 and HSP70 in 293 T cells overexpressing ASB10-FLAG and HSPA1-Myc. N = 3. E Co-immunoprecipitation assay to validate the interaction between HSP70 and ASB10 in 293 T cells overexpressing HSPA1-Myc and ASB10-FLAG. N = 3. F Representative images of anti-Asb10 and anti-HSP70 immunofluorescence staining. The right panel showed the colocalization of Asb10 and HSP70. PCCs r value was shown to quantify the colocalization. Scale bar = 10 μm. N = 5. G Schematic illustration of the structure of HSPA1, ASB10 and truncated HSPA1, ASB10. H Co-immunoprecipitation assay to detect the binding domain of HSPA1 with ASB10 in 293 T cells overexpressing the full length of HSPA1 and truncated ASB10. N = 3. I Co-immunoprecipitation assay to detect the binding domain of ASB10 with HSPA1 in 293 T cells overexpressing the full length of ASB10 and truncated HSPA1. N = 3.

Article Snippet: Antibody against Asb10 (#sc-100677) was purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Immunoprecipitation, Mass Spectrometry, Co-Immunoprecipitation Assay, Immunofluorescence, Staining, Binding Assay

Journal: Cell Death & Disease

Article Title: Asb10 accelerates pathological cardiac remodeling by stabilizing HSP70

doi: 10.1038/s41419-025-07735-5

Figure Lengend Snippet: A Protein expression of HSP70 after overexpressing Asb10 in NRVMs as determined by western blot. The right panel showed the quantifications. N = 6. B Protein expression of HSP70 after overexpressing ASB10 in 293 T cells as determined by western blot. The right panel showed the quantifications. N = 6. C Relative expressions of Asb10 and Hspa1 in NRVMs overexpressing Asb10 as determined by qPCR. N = 5. D Expression changes of HSP70 in NRVMs overexpressing Asb10 and treated with different time courses of CHX. The right panel showed the quantifications. N = 3. E Expression changes of HSP70 in 293T cells overexpressing ASB10 and treated with different time courses of CHX. The right panel showed the quantifications. N = 3. F Expression changes of HSP70 in NRVMs overexpressing Asb10 and treated with CQ or MG132 for 6 h. The right panel showed the quantifications. N = 3. G Co-immunoprecipitation assay to detect the changes of the STUB1/HSP70 complex and HSP70 ubiquitination level in NRVMs overexpressing Asb10. N = 3. H Co-immunoprecipitation assay to detect the changes of the STUB1/HSP70 complex and HSP70 ubiquitination level in NRVMs transfected with STUB1 siRNA and Ad- Asb10 . N = 3. I Co-immunoprecipitation assay to detect the changes of the STUB1/HSP70 complex and HSP70 ubiquitination level in 293T cells transfected with STUB1-EGFP, ASB10-FLAG, HSPA1-Myc and Ub-HA. N = 3. Unpaired Student’s t -test ( A – E ) was conducted. One-way ANOVA followed by Turkey’s multiple comparisons test ( F ) was conducted. Data were represented as Mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns P > 0.05.

Article Snippet: Antibody against Asb10 (#sc-100677) was purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Expressing, Western Blot, Co-Immunoprecipitation Assay, Ubiquitin Proteomics, Transfection

Journal: Cell Death & Disease

Article Title: Asb10 accelerates pathological cardiac remodeling by stabilizing HSP70

doi: 10.1038/s41419-025-07735-5

Figure Lengend Snippet: NRVMs were infected with adenovirus-packed Asb10 or Vector, and co-transfected with siNC or siHSP70 for 30 h, then treated with 50 μM PE for 24 h, and cells were then collected for further detection. A Protein expression of hypertrophic markers with GAPDH as loading control as determined by western blot. The right panel showed the quantifications. N = 6. B Relative mRNA expression of hypertrophic markers as detected by qPCR. N = 6. C Representative images of anti-α-actinin immunofluorescence staining showing cell size. The lower panel showed the quantifications. Scale bar = 30 μm. N = 76 (group 1), 79 (group 2), 82 (group 3), 84 (group 4), 75 (group 5) and 80 (group 6). Three-way ANOVA followed by Turkey’s multiple comparisons test was conducted. Data were represented as Mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns P > 0.05.

Article Snippet: Antibody against Asb10 (#sc-100677) was purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Infection, Plasmid Preparation, Transfection, Expressing, Control, Western Blot, Immunofluorescence, Staining

Journal: Cell Death & Disease

Article Title: Asb10 accelerates pathological cardiac remodeling by stabilizing HSP70

doi: 10.1038/s41419-025-07735-5

Figure Lengend Snippet: Mice were randomly divided into 4 groups: AAV9- EV +Sham, AAV9- Asb10 +Sham, AAV9- EV + TAC and AAV9- Asb10 + TAC. AAV9 infection was achieved via tail vein injection. Four weeks after injection, mice received either TAC or sham surgery. The cardiac function of mice was assessed using a non-invasive high-resolution imaging system. Mice were then sacrificed, and left ventricular tissues were collected for further detection. A Representative color mode echo images of blood flow in the aortic arch area (upper panel) and M mode images of echocardiography from 4 groups (lower panel). N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). B Ejection fraction (EF%) of mice in the late phase of TAC. N = 7 ( EV + Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). C Representative visual images of hearts from 4 groups. Scale bar = 1 cm. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). D Heart weight and body weight ratio (HW/BW). N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). E Representative H&E staining images of mice. Scale bar = 1 mm. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). F Representative WGA immunofluorescence staining images showing cardiomyocyte cross-sectional area. The right panel showed the quantifications. Scale bar = 200 μm. N = 216 ( EV +Sham), 180 ( Asb10 +Sham), 240 ( EV + TAC), 255 ( Asb10 + TAC). G Representative Masson’s and Sirius Red images showing cardiac interstitial fibrosis. The right panel showed the quantifications. Scale bar = 200 μm. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). H Protein expression of hypertrophic and fibrotic markers with GAPDH as loading control as determined by western blot. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). I Relative mRNA expression of pathological hypertrophic markers (upper panel) and cardiac fibrotic markers (lower panel) as detected by qPCR. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). Two-way ANOVA followed by Turkey’s multiple comparisons test was conducted. Data were represented as Mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Antibody against Asb10 (#sc-100677) was purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Infection, Injection, Imaging, Staining, Immunofluorescence, Expressing, Control, Western Blot

Journal: Cell Death & Disease

Article Title: Asb10 accelerates pathological cardiac remodeling by stabilizing HSP70

doi: 10.1038/s41419-025-07735-5

Figure Lengend Snippet: A ELISA assay to detect serum HSP70 levels. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). B Relative mRNA expression of Hspa1 and Asb10 as detected by qPCR. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). C Relative mRNA expression of canonical inflammation markers as detected by qPCR. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). D Representative immunohistochemistry staining images showing cardiac inflammation of mouse hearts. The right panel showed the quantifications. Scale bar = 200 μm. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). E Protein expression of pHDAC2 S394 , HSP70 and HDAC2 in left ventricular tissue of mice, with GAPDH as loading control as determined by western blot. The right panel showed the quantifications. N = 7 ( EV +Sham, Asb10 +Sham, EV + TAC) and 10 ( Asb10 + TAC). F Protein expression of pHDAC2 S394 , HSP70 and HDAC2 in Asb10 OE and HSP70 KD NRVMs treated with PE, with GAPDH as loading control as determined by western blot. The right panel showed the quantifications. N = 6. Two-way ANOVA followed by Turkey’s multiple comparisons test ( A – E ) was conducted. Three-way ANOVA followed by Turkey’s multiple comparisons test ( F ) was conducted. Data were represented as Mean ± SEM. * P < 0.05, ** P < 0.01, **** P < 0.0001, ns P > 0.05.

Article Snippet: Antibody against Asb10 (#sc-100677) was purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemistry, Staining, Control, Western Blot