Journal: Journal of Advanced Research

Article Title: Discovery of FBP1 as novel therapeutic target and asiatic acid-hydrogen sulfide donors accelerate diabetic wound healing

doi: 10.1016/j.jare.2025.12.003

Figure Lengend Snippet: Screening-identified asiatic acid inhibits FBP1 and promotes wound healing. (A) The relative FBP1 activity after the candidate compounds (50 μM) treatment using the FBP1 activity assay kit. (B) The chemical structure of AA. (C) Molecular docking structural model depicting asiatic acid-FBP1 binding interactions at the catalytic pocket (D) The effect of AA and AMP (0.01–100 μM) on FBP1 activity was detected by FBP1 activity assay kit. (E) CETSA analysis demonstrated enhanced FBP1 thermostability in asiatic acid-treated HaCaT cells. (F) Scratch wound healing assay demonstrated the migration changes after AA administration in HaCaT cells stimulated with MGO and overexpressing FBP1. (G) HaCaT cells exposed to asiatic acid (0.2–200 μM) for 24 h underwent MTT assay to quantify dose-dependent viability effects. (H) Clonal numbers after AA administration in HaCaT cells stimulated with MGO and overexpressing FBP1. (I) Transwell assay assessing invasion of HaCaT cells stimulated with MGO and overexpressing FBP1 after the introduction of AA. (J, K) Observation of HaCaT cells proliferation following AA administration using Ki67 staining (Scale bar = 100 μm). (L) Phalloidin staining after AA introduction in HaCaT cells stimulated with MGO and overexpressing FBP1. Data represent mean ± SEM from three independent replicates. * P < 0.05, ** P < 0.01, *** P < 0.001 vs Ctrl group. ^ P < 0.05, ^^ P < 0.01 vs OE group. & P < 0.05, && P < 0.01 vs NC group. # P < 0.05, ## P < 0.01 vs MGO group.

Article Snippet: For immunostaining, primary antibody targeting Ki67 (ABclonal, Wuhan, China) was applied and incubated overnight at 4 °C.

Techniques: Activity Assay, Binding Assay, Wound Healing Assay, Migration, MTT Assay, Transwell Assay, Staining

Journal: iScience

Article Title: Uhrf1 in PDGFRα-lineage cells regulates osteophyte formation in osteoarthritis

doi: 10.1016/j.isci.2026.116927

Figure Lengend Snippet: Uhrf1-positive cells are localized within the synovial thickening, and both Uhrf1 and PDGFRα double-positive cells are present within the mouse knee synovium (A) Uhrf1-positive cell localization within the osteophyte 4 days, 1 week, and 2 weeks after DMM in wild-type (WT) mice. The images show Safranin O/fast green and immunohistochemical staining against DAPI (blue), Uhrf1 (green), and KI67 (red). The borders between the bone and osteophyte (white solid line), between the chondrogenic area and synovial hypertrophic area (white dotted line), the hypertrophic synovial (HS) area (red arrow), and the chondrogenic area (CA) (blue arrow) are shown. Scale bars: 100 μm. (B) The images show Safranin O/fast green and immunohistochemistry for DAPI (blue), Uhrf1 (green), and PDGFRα (magenta) on knee synovium of WT mice 2 weeks after DMM. Uhrf1 and PDGFRα double-positive cells are designated with white arrows. Scale bars: 200 μm.

Article Snippet: Immunofluorescence staining was performed on tissue specimens by autoclaving with 0.5% ImmunoSaver (Wako, Osaka, Japan) for 45 min at 85°C and blocking by treatment with a solution of 1% BSA and 0.02% Triton in PBS for 1 h. Primary antibodies were mouse anti-Uhrf1 monoclonal antibody (Santa Cruz Biotechnology, TX, USA; 1:400), rat anti-KI67 monoclonal antibody (Synaptic systems, Goettingen, Germany; 1:400), goat anti-PDGFRα monoclonal antibody (R&D Systems, MSP, USA; 1:400), rat anti-GFP monoclonal antibody (R&D Systems, MSP, USA; 1:400).

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry

Journal: iScience

Article Title: Uhrf1 in PDGFRα-lineage cells regulates osteophyte formation in osteoarthritis

doi: 10.1016/j.isci.2026.116927

Figure Lengend Snippet: PDGFRα-lineage cell-specific Uhrf1-knockout mice showed reduced osteophyte formation 2 weeks after DMM (A) Uhrf1 mRNA expression levels in PDGFRα-positive cells obtained from lower limb muscles of Ctrl and cKO mice at 12 weeks of age, using FACS. Biological replicates, n = 4. The dots show individual data points. All data are mean ± SD. ∗ p < 0.05 (unpaired two-tailed Student’s t test). (B) Immunocytochemistry for DAPI (blue), Uhrf1 (red), and KI67 (green) on PDGFRα-positive cells from lower limb muscles of Ctrl and cKO mice at 12 weeks of age, using FACS. Scale bars: 20 μm. (C) Uhrf1-positive proportion of PDGFRα-positive cells. Biological replicates, n = 3. The dots show individual data points. All data are mean ± SD. ∗∗∗ p < 0.001 (Mann-Whitney U test). (D) Osteophytes at the medial tibial articular surface with lower (left) and higher (right) magnifications of Ctrl and cKO mice 2 weeks after DMM, stained with Safranin O and fast green. Scale bars: 200 μm. (E–G) The upper line graphs show (E) osteophyte width, (F) osteophyte area, and (G) modified maturity score at each evaluation site for Ctrl and cKO mice 2 weeks after DMM. The dots show the mean values. The lower graphs show area under the curve (AUC) for the osteophyte width, osteophyte area, and modified maturity score. Biological replicates, n = 9. The dots show individual data points. All data are mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01 (Mann-Whitney U test). DMM, destabilization of the medial meniscus.

Article Snippet: Immunofluorescence staining was performed on tissue specimens by autoclaving with 0.5% ImmunoSaver (Wako, Osaka, Japan) for 45 min at 85°C and blocking by treatment with a solution of 1% BSA and 0.02% Triton in PBS for 1 h. Primary antibodies were mouse anti-Uhrf1 monoclonal antibody (Santa Cruz Biotechnology, TX, USA; 1:400), rat anti-KI67 monoclonal antibody (Synaptic systems, Goettingen, Germany; 1:400), goat anti-PDGFRα monoclonal antibody (R&D Systems, MSP, USA; 1:400), rat anti-GFP monoclonal antibody (R&D Systems, MSP, USA; 1:400).

Techniques: Knock-Out, Expressing, Muscles, Two Tailed Test, Immunocytochemistry, MANN-WHITNEY, Staining, Modification

Journal: mBio

Article Title: Potential mechanisms underlying Enterococcus faecalis -driven pancreatic cancer cell proliferation

doi: 10.1128/mbio.03963-25

Figure Lengend Snippet: E. faecalis promoted the proliferation of pancreatic cancer cells. ( A ) An in vitro co-culture system was established to explore the influence of E. faecalis on pancreatic cancer cells. PANC1 and PANC02 cells were briefly co-cultured with V583 or EF at various MOIs, followed by removal of extracellular bacteria via washing and further culture for 48 h with a triple-antibiotic cocktail. ( B ) Cell survival was assessed by Calcein-AM/PI staining after 4 h of co-culture. Scale bar = 100 µm. ( C ) The impact of E. faecalis on the viability of pancreatic cancer cells was assessed using MTT assay at 48 h. ( D and E ) PANC1 or PANC02 cell numbers were quantified using a cell counter after 48 h with or without V583 stimulation. ( F ) The proliferation of PANC1 cells was assessed by EdU assay at multiple time points post-V583 infection. ( G and H ) Ki67 immunofluorescence staining and quantitative analysis were performed 48 h after infection with V583 ( G ) or EF ( H ) to assess PANC1 proliferation. * P < 0.05, ** P < 0.01 ( N = 6).

Article Snippet: Endogenous peroxidase activity was blocked with serum, and the cells were incubated overnight at 4°C with anti-Ki67 primary antibody (Abclonal, 1:1000).

Techniques: In Vitro, Co-Culture Assay, Cell Culture, Bacteria, Staining, MTT Assay, EdU Assay, Infection, Immunofluorescence

Journal: mBio

Article Title: Potential mechanisms underlying Enterococcus faecalis -driven pancreatic cancer cell proliferation

doi: 10.1128/mbio.03963-25

Figure Lengend Snippet: E. faecalis enhanced the migration and invasion of pancreatic cancer cells. ( A ) Wound healing assay showing PANC1 cell migration after V583 infection (scale bar = 200 µm). ( B ) Wound healing assay assessing the effect of EF infection on PANC1 cell migration (scale bar = 200 µm). ( C, D ) Quantification of wound closure in panels A and B . ( E ) Transwell assay evaluating the invasive potential of PANC1 cells after V583 infection (scale bar = 100 µm). ( F ) Statistical analysis of panel E . ( G ) Transwell assay assessing invasion of EF-infected PANC1 cells (scale bar = 100 µm). ( H ) Statistical analysis of panel G . ( I ) Western blot analysis of proliferation markers (PCNA, cyclin D1) and apoptosis markers (caspase-9, cleaved caspase-3) in PANC1 cells following V583 stimulation. ( J ) Quantitative analysis of panel I . ( K ) RNA-seq data showing upregulation of proliferation-related genes (CCNB2, CCND1, PCNA, MKI67) in PANC1 cells 48 h post-V583 treatment. ( L ) Western blot analysis of E-cadherin and N-cadherin expression in V583-treated PANC1 cells. ( M ) Quantitation of panel L . ( N ) RNA-seq results showing altered expression of E-cadherin and N-cadherin genes in PANC1 cells after V583 stimulation. * P < 0.05, ** P < 0.01 ( N = 4).

Article Snippet: Endogenous peroxidase activity was blocked with serum, and the cells were incubated overnight at 4°C with anti-Ki67 primary antibody (Abclonal, 1:1000).

Techniques: Migration, Wound Healing Assay, Infection, Transwell Assay, Western Blot, RNA Sequencing, Expressing, Quantitation Assay

Journal: mBio

Article Title: Potential mechanisms underlying Enterococcus faecalis -driven pancreatic cancer cell proliferation

doi: 10.1128/mbio.03963-25

Figure Lengend Snippet: The impact of E. faecalis on pancreatic tumor growth in vivo . ( A ) Schematic of the in vivo experimental model for V583 infection in pancreatic cancer. ( B ) Representative ex vivo tumor images at day 60, showing visible differences between V583 and control groups. ( C ) Comparison of tumor weights between the V583-inoculated and control groups at day 60. * P < 0.05 ( N = 6). ( D ) H&E staining and immunohistochemical analysis of EGFR and Ki67 expression in tumor tissues.

Article Snippet: Endogenous peroxidase activity was blocked with serum, and the cells were incubated overnight at 4°C with anti-Ki67 primary antibody (Abclonal, 1:1000).

Techniques: In Vivo, Infection, Ex Vivo, Control, Comparison, Staining, Immunohistochemical staining, Expressing