anti mouse cd38 pe vio770  (Miltenyi Biotec)

Journal: Molecular Biomedicine

Article Title: EZH2 coordinates inflammatory and pain-associated pathways across joint compartments in osteoarthritis

doi: 10.1186/s43556-026-00643-w

Figure Lengend Snippet: EZH2 inhibition reduced the inflammatory phenotype of macrophages during OA. Macrophage differentiation and M1-like polarization were induced from the THP-1 monocytic cell line ( a ) or from human bone marrow-derived cells isolated from OA patients ( b ) in the presence or absence of EPZ-6438. The expression of pro-inflammatory markers (IL-1β, IL-6 and CXCL1) was analyzed by RT-qPCR. Experiments using THP-1 cells were independently repeated at least three times. Experiments using human bone marrow-derived cells were performed using biological replicates from independent OA patients. Data are presented as mean ± SEM. *: p -value < 0.05; **: p -value < 0.01; ***: p -value < 0.001. Spleens from OA mice described in Fig. and treated with vehicle or EPZ-6438 were collected and analyzed by flow cytometry. Cells were stained for macrophage-associated markers ( c ). Samples were acquired using a FACSVerse flow cytometer and analyzed with FlowJo 7.6.5 software. Results are presented as median ± interquartile range (n = 8 mice). *: p -value < 0.05; **: p -value < 0.01

Article Snippet: The following antibodies were used for immune cell characterization: CD11b (REA592 VioBlue, 130–113–810, Miltenyi Biotec), F4/80 (REA126 PE-Vio770, 130–118–459, Miltenyi Biotec), CD80 (REA983 APC-Vio770, 130–116–463, Miltenyi Biotec), CD86 (REA1190 PE, 130–122–129, Miltenyi Biotec), MHC-II (REA813 FITC, 130–112–386, Miltenyi Biotec), CD38 (REA616 PerCP-Vio700, 130–109–260, Miltenyi Biotec), and iNOS (REA982 APC, 130–116–423, Miltenyi Biotec).

Techniques: Inhibition, Derivative Assay, Isolation, Expressing, Quantitative RT-PCR, Flow Cytometry, Staining, Software

Journal: Molecular Biomedicine

Article Title: EZH2 coordinates inflammatory and pain-associated pathways across joint compartments in osteoarthritis

doi: 10.1186/s43556-026-00643-w

Figure Lengend Snippet: Proposed model illustrating the effects of EZH2 inhibition across multiple joint compartments in osteoarthritis. In OA fibroblast-like synoviocytes (FLS), increased EZH2 and H3K27me3 levels were associated with inflammatory and catabolic responses, which were reduced by pharmacological EZH2 inhibition with EPZ-6438. In human OA FLS, EPZ-6438 decreased inflammatory, catabolic and pain-associated mediators, including IL1B, MMP13, NGF and NTN1, reduced LDHA expression and increased autophagy-associated responses. Proteomic and ChIP-seq analyses further identified inflammatory, metabolic and axon guidance-related pathways, including inflammation-dependent EZH2 recruitment at the PAX6 locus. EZH2 inhibition also reduced inflammatory activation of macrophages and osteoclastogenesis in OA-derived bone marrow cells. Collectively, these findings support a broader role for EZH2 in inflammatory, catabolic and osteoimmune responses across multiple OA-relevant cellular compartments. The identification of PAX6, NTN1 and axon guidance-related pathways highlights neuro-associated molecular programs that may be relevant to pain-associated responses, although their direct contribution to pain or sensory nerve remodeling was not assessed in this study. The reduced structural joint damage and pain-related behaviors observed following EZH2 inhibition in the MIA-induced OA model are consistent with this multicellular framework

Article Snippet: The following antibodies were used for immune cell characterization: CD11b (REA592 VioBlue, 130–113–810, Miltenyi Biotec), F4/80 (REA126 PE-Vio770, 130–118–459, Miltenyi Biotec), CD80 (REA983 APC-Vio770, 130–116–463, Miltenyi Biotec), CD86 (REA1190 PE, 130–122–129, Miltenyi Biotec), MHC-II (REA813 FITC, 130–112–386, Miltenyi Biotec), CD38 (REA616 PerCP-Vio700, 130–109–260, Miltenyi Biotec), and iNOS (REA982 APC, 130–116–423, Miltenyi Biotec).

Techniques: Inhibition, Expressing, ChIP-sequencing, Activation Assay, Derivative Assay

Journal: Advanced Science

Article Title: Bone Marrow Macrophage STING Signaling is a Direct Nexus Linking Marrow Adipose Expansion to NAD + Depletion and Osteogenic Decline in Skeletal Aging

doi: 10.1002/advs.77571

Figure Lengend Snippet: Enhanced CD38 ecto‐NADase activity in BMMs during skeletal aging impairs NMN availability to BMSCs. (a) qPCR analysis of Cd38 , Cdkn2a , Cdkn1a , Nos2 , Tnfα , IL1b , IL6 , Ym1 , Fizz1 , and Arg1 mRNA levels in bone marrow from 3‐ or 18‐month‐old WT and 18‐month‐old Sting ΔMφ mice ( n = 6 biological replicates). (b—d) Representative micro‐CT images (b) and quantitative analysis of BV/TV, Tb. N, Tb.Th, and Tb.Sp (c,d) from AAV9‐BMM‐shNC‐treated or AAV9‐BMM‐sh Cd38 ‐treated male mice ( n = 6 biological replicates). (e) WST‐8 analysis of NAD + levels in 3, 10, 18, and 24 M‐BMSCs ( n = 6 biological replicates). (f) WST‐8 analysis of NAD + levels in BMSCs under co‐culture conditions ( n = 6 biological replicates). (g) WST‐8 analysis of NAD + levels in BMSCs from the bone marrow of mice treated with PBS or 78c from 16‐ to 18‐month‐old ( n = 6 biological replicates). (h) WST‐8 analysis of NAD + levels in BMSCs from the bone marrow of mice injected with AAV9 for BMM‐specific overexpression of WT or Δ49 mouse Cd38 ( n = 6 biological replicates). Data are shown as mean ± SD.

Article Snippet: In parallel, 16‐month‐old male C57BL/6J mice received 78c (MedChemExpress, HY‐123999; 10 mg/kg) or PBS intraperitoneally 5 days per week for 8 weeks.

Techniques: Activity Assay, Analysis, Micro-CT, Co-culture Assay, Injection, Over Expression