Fig. 5 | Bone Research

Fig. 5

From: Akkermansia muciniphila attenuates intervertebral disc degeneration via extracellular vesicle-mediated delivery of the effector protein B2UKX5

Fig. 5The alternative text for this image may have been generated using AI.

B2UKX5 as a potential functional protein in Akk-EVs against IVDD. Hierarchical clustering heatmap (a) and count (b) of differentially enriched proteins between Akk and Akk-EVs. c Functional annotation for the highly enriched 388 proteins using the COG database. d Relative enrichment levels of the five selected candidate proteins in Akk-EVs compared to Akk, displayed as Akk-EVs/Akk ratios. e Heatmap showing qPCR-based expression levels of MMP13, ADAMTS5, TNF, and CDKN1A in TNF-α-treated human NP cells following treatment with the five candidate proteins. n = 3 per group. f Cytotoxicity assessment of B2UKX5 in human NP cells using Cell Counting Kit-8 assay after 24 h treatment under indicated concentrations. Cell viability is expressed relative to vehicle control. n = 3 per group. g Representative IF staining of COL2 (green) in TNF-α-treated NP cells after B2UKX5 intervention; Nuclei are counterstained with DAPI (blue). Scale bar: 100 μm. h Quantification of COL2 mean fluorescence intensity in human NP cells. n = 3 per group. i Representative IF staining of MMP13 (green) in TNF-α-treated NP cells after B2UKX5 intervention; Nuclei are counterstained with DAPI (blue). Scale bar: 100 μm. j Quantification of MMP13 mean fluorescence intensity in human NP cells. n = 3 per group. k Correlation analysis between age, IVDD grading (Pfirrmann), and serum B2UKX5 levels in human subjects. n = 79. l Quantification of B2UKX5 levels in serum from mice of different age groups. n = 15–16 per group. m Representative IF staining of B2UKX5 (green) in human NP tissue from patients with varying degrees of IVDD; Nuclei are counterstained with DAPI (blue). Scale bar: 50 μm. n Quantification of B2UKX5-positive area (%) in human NP tissue; n = 9–11 per group. o Representative IF staining of B2UKX5 (green) in intervertebral disc sections from young versus aged mice; Nuclei are counterstained with DAPI (blue). Scale bar: 100 μm. p Quantification of B2UKX5-positive area (%) in mouse discs. n = 9–10 per group. Data are presented as mean ± SD. Statistical significance was assessed using one-way ANOVA followed by Bonferroni post hoc test, no significant differences were observed in panel f, and Pearson correlation was used for panel k

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