Journal: Neural Regeneration Research

Article Title: Porcine decellularized nerve matrix hydrogel attenuates neuroinflammation after peripheral nerve injury by inhibiting the TLR4/MyD88/NF-κB axis

doi: 10.4103/NRR.NRR-D-24-00302

Figure Lengend Snippet: TLR4/MyD88/NF-κB pathway plays an indispensable role in regulating pDNM-gel-mediated macrophage polarization and anti-inflammatory reactions in vitro . (A) WB detection of TLR4, MyD88, p-NF-κB, NF-κB, p-IκBα, and IκBα in RAW264.7 cells treated with LPS (100 ng/mL) + pDNM-gel (0.5%) for 24 hours, followed by the addition of CRX-527 (500 µM), TAK242 (100 nM), CL075 (2 µM), or T6167923 (100 µM) for another 24 hours. (B–E) Quantitative analysis of the expression levels of these four proteins from A. (F, G) Co-staining of CD68 (red) with iNOS (green) or Arg1 (green) in the LPS + pDNM-gel, LPS + pDNM-gel + CRX-527, LPS + pDNM-gel + TAK242, LPS + pDNM-gel + CL075, and LPS + pDNM-gel + T6167923 treatment groups. Nuclei were stained with DAPI (blue). Scale bars: 10 μm. (H, I) Quantification of the fluorescence intensity of iNOS and Arg1 from F and G. (J, K) Flow cytometry detection of M1-type and M2-type macrophages in each group. (L, M) Quantitative analysis of the percentages of double-positive CD68 + CD86 + and CD68 + CD206 + cells from J and K. (N, O) qRT-PCR analysis of the mRNA expression of pro-inflammatory signature genes TNF-α , IL-1 β, and IL-6 , and anti-inflammatory signature genes IL-4 , IL-10 , and TGF- β, in each group. (P, Q) ELISAs of inflammatory cytokine levels in RAW264.7 cell culture supernatants in each group. Data are expressed as mean ± SEM from three independent assays in vitro ; * P < 0.05, ** P < 0.01, *** P < 0.001. Arg1: Arginase 1; CD206: cluster of differentiation protein 206; CD68: cluster of differentiation protein 68; CD86: cluster of differentiation protein 86; DAPI: 4′,6-Diamidino-2-phenylindole; ELISA: Enzyme-linked immunosorbent assay; IL-10: interleukin-10; IL-1β: interleukin-1β; IL-4: interleukin-4; IL-6: interleukin-6; iNOS: inducible nitric oxide synthase; IκBα: nuclear factor kappa B inhibitory protein alpha; LPS: lipopolysaccharides; MyD88: myeloid differentiation factor 88; NF-κB: nuclear factor-κB; n.s,: not significant; pDNM-gel: porcine decellularized nerve matrix hydrogel; p-IκBα: phosphorylated nuclear factor kappa B inhibitory protein alpha; p-NF-κB: phosphorylated nuclear factor-κB; qRT-PCR: quantitative reverse transcription-polymerase chain reaction; TGF-β: transforming growth factor-beta; TLR4: Toll-like receptor 4; TNF-α: tumor necrosis factor-alpha.

Article Snippet: Then, the medium was added with one of the following agonists/inhibitors and treated the cells for a further 24 hours: 500 μM CRX-527 (InvivoGen, Toulouse, France, Cat# tlrl-crx527), 100 nM TAK242 (MedChemExpress, Monmouth Junction, NJ, USA, Cat# HY-11109), 2 μM CL075 (MedChemExpress, Cat# HY-117066), or 100 μM T6167923 (MedChemExpress, Cat# HY-19744; Additional Figure 3 ).

Techniques: In Vitro, Expressing, Staining, Fluorescence, Flow Cytometry, Quantitative RT-PCR, Cell Culture, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Porcine decellularized nerve matrix hydrogel attenuates neuroinflammation after peripheral nerve injury by inhibiting the TLR4/MyD88/NF-κB axis

doi: 10.4103/NRR.NRR-D-24-00302

Figure Lengend Snippet: Promotional effect of pDNM-gel on nerve regeneration is abolished using TLR4/MyD88/NF-κB agonists in vivo . (A) Immunofluorescence staining for neurofilament-H (green) and MBP (red) in sciatic nerves from rats treated with pDNM-gel, pDNM-gel + CRX-527, or pDNM-gel + CL075 at 21 dpi ( n = 5/group). Nuclei were stained with DAPI (blue). Scale bars: 20 μm. (B) Representative TEM images of regenerated sciatic nerve cross-sections from the three groups at 21 dpi. Scale bars: 10 μm. (C, D) Calculated area (%) of positive staining of neurofilament-H and MBP per 100 μm 2 in the immunofluorescence images for the three groups ( n = 5/group). (E, F) Statistical analysis of the number and thickness of myelin sheaths in the three groups using ImageJ software ( n = 3/group). Data are expressed as mean ± SEM; ** P < 0.01, *** P < 0.001. DAPI: 4′,6-Diamidino-2-phenylindole; dpi: days post-injury; MBP: myelin basic protein; MyD88: myeloid differentiation factor 88; NF-κB: nuclear factor-κB; pDNM-gel: porcine decellularized nerve matrix hydrogel; PNI: peripheral nerve injury; SEM: standard error of the mean; TEM: transmission electron microscopy; TLR4: Toll-like receptor 4.

Article Snippet: Then, the medium was added with one of the following agonists/inhibitors and treated the cells for a further 24 hours: 500 μM CRX-527 (InvivoGen, Toulouse, France, Cat# tlrl-crx527), 100 nM TAK242 (MedChemExpress, Monmouth Junction, NJ, USA, Cat# HY-11109), 2 μM CL075 (MedChemExpress, Cat# HY-117066), or 100 μM T6167923 (MedChemExpress, Cat# HY-19744; Additional Figure 3 ).

Techniques: In Vivo, Immunofluorescence, Staining, Software, Transmission Assay, Electron Microscopy

Journal: bioRxiv

Article Title: SLAMF1-peptide mediated epigenetic priming reprograms innate immune responses in sepsis

doi: 10.64898/2025.12.29.696918

Figure Lengend Snippet: Volcano plots comparing cytokine secretion by PBMCs primed with vehicle or P7-Pen and subsequently stimulated with LPS ( A ) or CL075 ( B ) for 2 h. Plots were generated by plotting log 2 fold change against −log 10 FDR-adjusted p values (q < 0.05). (C) Quantification of the most strongly affected LPS-induced cytokines in PBMCs primed with vehicle or P7-Pen. Statistical significance for panels (A–C) was assessed using multiple unpaired t -tests ( p < 0.05, * p < 0.01, ** p < 0.001).

Article Snippet: Ultrapure K12 LPS (#tlrl-peklps) from E. coli , ultrapure thiazoloquinoline compound CL075 (#tlrl-c75), benzazepine analog TL8-506 (#tlrl-tl8506) and FSL-1 TLR2/TLR6 ligand (#tlrl-fsl) were from InvivoGen (San Diego, CA, USA).

Techniques: Generated

Journal: bioRxiv

Article Title: SLAMF1-peptide mediated epigenetic priming reprograms innate immune responses in sepsis

doi: 10.64898/2025.12.29.696918

Figure Lengend Snippet: Western blot analysis shows histone H3 acetylation patterns together with quantitative densitometry for all analyzed samples. (A, B) Representative immunoblots of PBMC lysates from healthy donors ( A ) or sepsis patients ( B ) showing levels of H3K9ac, H3K27ac, and H3K56ac following priming with vehicle (water), 15 µM control peptide C3-Pen, or 15 µM P7-Pen for 20–22 h. (C) Representative immunoblots (2 of 10 donors per group) showing H3K27ac levels in PBMCs primed with water or P7-Pen and subsequently stimulated with LPS or CL075 for 2 h. Quantification shown in the graphs represents signal intensity normalized to the corresponding loading control (β-tubulin). Data are presented as mean relative fold change ± SEM. Statistical significance was assessed using the nonparametric Wilcoxon matched-pairs signed-rank test ( p < 0.05, * p < 0.01, ** p < 0.001).

Article Snippet: Ultrapure K12 LPS (#tlrl-peklps) from E. coli , ultrapure thiazoloquinoline compound CL075 (#tlrl-c75), benzazepine analog TL8-506 (#tlrl-tl8506) and FSL-1 TLR2/TLR6 ligand (#tlrl-fsl) were from InvivoGen (San Diego, CA, USA).

Techniques: Western Blot, Control