Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: Targeting Capability of D-EVs to Senescent NPCs is Mediated by the CXCL10-CXCR3 Axis. (A) Schematic diagram of RNA sequencing for senescent NPCs treated with D-EVs or not. (B-C) Volcano plots and heatmaps displaying differential gene expression in senescent NPCs treated with D-EVs or not. (D) A Venn diagram illustrates overlap between DEGs identified in senescent NPCs treated with D-EVs and key gene databases for cytokine-cytokine receptor interactions. (E) A Venn diagram illustrates overlap between DEGs identified in D-MSCs and chemokine signaling pathway key gene databases. (F-G) Network analysis of the hub genes among the above total DEGs. A protein-protein interaction network was created in the STRING database, while the Cytoscape software was used to determine the hub genes in the network. (H) GO analysis confirming enrichment of terms related to senescent NPCs treated with D-EVs or not in the BP categories. (I) GSEA analysis of cytokine-cytokine receptor interactions in D-EVs group versus TBHP group. (J) Western blot analysis confirmed the expression of CXCR3 in Control and senescent NPCs, and (S) quantitative analysis. (K) Western blot analysis confirmed the expression of CXCL10 in N-EVs and D-EVs, and (R) quantitative analysis. (L) Protein binding analysis of CXCR3 and CXCL10 by PyMOL. (M) Confocal analysis of the uptake of pHrodo-labeled EVs in senescent NPCs following the treatments of N-EVs, D-EVs with CXCL10 knockout/anti-CXCR3, or not, and (Q) quantitative analysis. (N) In vivo tracking of PKH26-labeled CXCL10 knockout D-EVs. (O) Flow cytometry showing uptake of different EVs with CXCL10 knockout/anti-CXCR3 or not by senescent NPCs, and (P) quantitative analysis. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membranes were incubated with primary antibodies against GAPDH (1:5000, 104941-AP, Proteintech), TSG101 (1:1000, DF8427, Affinity), CD9 (1:1000, AF5139, Affinity), CD63 (1:2000, 25682-1-AP, Proteintech), Calnexin (1:5000, 10427-2-AP, Proteintech), GM130 (1:20000, 11308-1-AP, Proteintech), CXCR3 (1:5000, 26756-1-AP, Proteintech), CXCL10 (1:2000, 10937-1-AP, Proteintech), MMP3 (1:2000, 17873-1-AP, Proteintech), ADAMTS5 (DF13268, Affinity), P16 (AF5484, Affinity), P21 (10355-1-AP, Proteintech), GPX4 (1:1000, 381958, Zen-bio), SLC7A11 (1:1000, 26864-1-AP, Proteintech), ACSL4 (1:5000, 22401-1-AP, Proteintech) and Tubulin (1:10000, T40103 , Abmart) overnight at 4 °C.

Techniques: RNA Sequencing, Gene Expression, Software, Western Blot, Expressing, Control, Protein Binding, Labeling, Knock-Out, In Vivo, Flow Cytometry

Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: D-EVs Deliver GPX4 to Inhibit Ferroptosis in Senescent NPCs. (A) Representative Senescent-Tracker images of NPCs treated with N-EVs, D-EVs, Era, and D-Evs sh-CXCL10 . (B) Volcano plot of transcriptomic data comparing D-MSC and N-MSC. (C) KEGG pathway analysis of DEGs in D-MSCs versus N-MSCs. (D) Volcano plot of proteomic data comparing D-EVs and N-EVs. (E) KEGG pathway analysis of transcriptomic and proteomic data integration. (F) A Venn diagram illustrating the intersection of genes from the D-MSC transcriptome, the D-EVs proteome, and the ferroptosis-related gene set. (G) Bar graph showing the relative expression levels of core overlapping genes identified in (F). (H) MS analysis revealed that GPX4 is enriched in the D-EVs proteome. (I) Western blot analysis confirming GPX4 protein in D-EVs and N-EVs. (J) Western blot analysis of key senescence (p21, P16) markers in NPCs following treatment with PBS or N-EVs with CXCL10 or GPX4 knockout. (K) Representative images of EdU depicting cell proliferation ability in the control, TBHP, D-EVs, D-EVs sh-CXCL10 , D-EVs sh-GPX4 , and D-EVs sh-CXCL10+GPX4 groups. (L-M) Confocal images showing GPX4 delivery from different EVs to senescent NPCs at 12h and 24h co-culture, and (N) colocalization analysis. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membranes were incubated with primary antibodies against GAPDH (1:5000, 104941-AP, Proteintech), TSG101 (1:1000, DF8427, Affinity), CD9 (1:1000, AF5139, Affinity), CD63 (1:2000, 25682-1-AP, Proteintech), Calnexin (1:5000, 10427-2-AP, Proteintech), GM130 (1:20000, 11308-1-AP, Proteintech), CXCR3 (1:5000, 26756-1-AP, Proteintech), CXCL10 (1:2000, 10937-1-AP, Proteintech), MMP3 (1:2000, 17873-1-AP, Proteintech), ADAMTS5 (DF13268, Affinity), P16 (AF5484, Affinity), P21 (10355-1-AP, Proteintech), GPX4 (1:1000, 381958, Zen-bio), SLC7A11 (1:1000, 26864-1-AP, Proteintech), ACSL4 (1:5000, 22401-1-AP, Proteintech) and Tubulin (1:10000, T40103 , Abmart) overnight at 4 °C.

Techniques: Expressing, Western Blot, Knock-Out, Control, Co-Culture Assay

Journal: bioRxiv

Article Title: CXCL10 drives female-specific tau pathology progression and defines sex-dependent vulnerability in tauopathy model mice

doi: 10.64898/2026.04.19.719088

Figure Lengend Snippet: A Schematic of the experimental workflow. Hippocampi and cerebral cortices were collected from 12-month-old Tau Tg mice and age-matched WT controls. Bulk RNA-seq was performed on the hippocampi, while protein array analysis and CXCL10 ELISA were conducted using the cerebral cortices. At this age, Tau Tg mice exhibit prominent AT8-positive tau phosphorylation and brain atrophy. b–e Transcriptomic analysis of hippocampal bulk RNA-seq from Tau Tg and WT mice (n = 3 per group). b PCA plot. c Heatmap of the top 1,000 DEGs. d GSEA highlighting immune-related pathways enriched in Tau Tg mice. e Volcano plot of DEGs in Tau Tg mice compared to WT controls. DEG criteria: log₂ fold change > |1| and p < 0.1. f, g Cytokine and chemokine expression in the cerebral cortices of 9-month-old Tau Tg mice compared with age-matched WT controls. f Representative images of a cytokine/chemokine array using tissue lysates from the cerebral cortex. g Quantification of relative expression levels of indicated proteins in Tau Tg mice. Data are presented as mean ± S.E.M. h, i Expression levels of CXCL10 protein in the cerebral cortex ( h ) and the hippocampus ( i ) were measured by ELISA. Red dots indicate female mice, and white dots indicate male mice Data are presented as mean ± S.E.M. Number of mice used: except for 11–12-month-old female Tau Tg mice, where seven mice were used, we used five mice for each condition. For b – e , DEGs were identified using the Ward test. Statistical analysis was performed using a mixed-effects analysis followed by a Šídák’s multiple comparisons test ( h, i ). * p < 0.05, **** p < 0.0001. Source data are provided in the Source Data file.

Article Snippet: B6.129S4- Cxcl10 tm1Adl /J ( Cxcl10 KO, RRID:IMSR_JAX:006087) mice and B6.129X1- Mapt tm1Hnd /J ( Mapt KO, RRID:IMSR_JAX:007251) mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: RNA Sequencing, Protein Array, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Expressing

Journal: bioRxiv

Article Title: CXCL10 drives female-specific tau pathology progression and defines sex-dependent vulnerability in tauopathy model mice

doi: 10.64898/2026.04.19.719088

Figure Lengend Snippet: Immunoblot analyses of tau protein in the cerebral cortices of 9-month-old Tau Tg mice with or without Cxcl10 deficiency. Males ( a–f ), females ( g–l ). a–e, g–k Immunoblot images and relative expression of phosphorylated tau (p-tau) detected with anti-pSer396, pThr212/pSer214 [AT100], and pSer202/pThr205 [AT8] antibodies, and total tau detected with TAU-5 antibody. Protein expression was normalized to total protein using the Stain-Free method. f, l Immunoblot images of sarkosyl-insoluble tau from the cerebral cortex, including the piriform and entorhinal cortices. Bar graphs represent relative expression levels of tau protein in Tau Tg; Cxcl10 KO mice compared to Tau Tg mice. m , o Immunohistochemical analysis of p-tau in the hippocampus using AT8 antibody. Bar graphs indicate the AT8-positive area in male ( n ) and female ( p ) mice. Scale bar = 500 µm. Data are presented as mean ± S.E.M. Number of mice used: Tau Tg (n = 5), and Tau Tg; Cxcl10 KO (n = 5) for both sexes. q , r Graphs show survival curve in Tau Tg and Tau Tg; Cxcl10 KO mice in males ( q ) and females ( r ). Number of mice used: Males: Tau Tg (n = 17) and Tau Tg; Cxcl10 KO (n = 20); Females: Tau Tg (n = 22) and Tau Tg; Cxcl10 KO (n = 21). Statistical analysis was performed using an unpaired two-tailed t-test ( b, d, e, f, h, l, n, p ), an unpaired t-test with Welch’s correction ( I, J, K ), a Mann-Whitney U test ( c ), and a log-rank test ( q , r ). * p < 0.05, ** p < 0.01. Source data, including full blot images are provided in the Source Data file.

Article Snippet: B6.129S4- Cxcl10 tm1Adl /J ( Cxcl10 KO, RRID:IMSR_JAX:006087) mice and B6.129X1- Mapt tm1Hnd /J ( Mapt KO, RRID:IMSR_JAX:007251) mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Western Blot, Expressing, Staining, Immunohistochemical staining, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: CXCL10 drives female-specific tau pathology progression and defines sex-dependent vulnerability in tauopathy model mice

doi: 10.64898/2026.04.19.719088

Figure Lengend Snippet: a Schematic diagram of the spatial transcriptome analysis using Xenium Prime 5K. Formalin fixed paraffin embedded (FFPE) sections were prepared from 12-month-old Tau Tg mice and age-matched WT controls. b UMAP visualizing the cell cluster detected by Xenium in the brains of Tau Tg and WT mice. Neuronal cells were classified as IT (intratelencephalic), ET (extratelencephalic), Glut (glutamatergic), NP (near-projecting), CT (corticothalamic), L6b (layer 6b), DG (dentate gyrus), IMN (immature neurons), CTX (cerebral cortex), CGE (caudal ganglionic eminence), GABA (GABAergic), MGE (medial ganglionic eminence), CNU (cerebral nuclei), LGE (lateral ganglionic eminence), Hya (anterior hypothalamic), HY (hypothalamus), MM (medial mammillary nucleus), LH (lateral habenula), TH (thalamus), MB (midbrain), HB (hindbrain), Sero (serotonergic), MY (medulla), NN (non-neuronal), NP (near-projecting), OB (olfactory bulb), OEC (olfactory ensheathing cells), and OLF (olfactory areas). c Cxcl10 mRNA signal was plotted using Feature Plot on UMAP. d Quantitative Cxcl10 gene expression using violin plots in AC-Epen, BAM, DG-IMN Glut, IT-ET Glut, MG, and T cell types. e, h Representative plots of the result of re-clustering AC-Epen ( e ) and immune cluster ( h ), respectively. f Plots of Cxcl10 + cells in the cluster shown in and represented according to genotype. g, j Figures showing spatial distribution of AC8 ( g ) and MG3 ( j ) clusters in the brains of WT and Tau Tg mice. k Representative images of coronal section of mouse brain by Xenium explorer. Scale bar = 1 mm. l Spatial information of Cxcl10 + astrocytes and microglia in the hippocampus of Tau Tg mice using Xenium explorer. Scale bar = 100 μm. Number of mice used: male WT (n = 1), male Tau Tg (n = 1), female WT (n = 1), and female Tau Tg (n = 1). Statistical analysis was performed using a Wilcoxon rank sum U statistic test ( d ). Source data are provided in the Source Data file.

Article Snippet: B6.129S4- Cxcl10 tm1Adl /J ( Cxcl10 KO, RRID:IMSR_JAX:006087) mice and B6.129X1- Mapt tm1Hnd /J ( Mapt KO, RRID:IMSR_JAX:007251) mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Formalin-fixed Paraffin-Embedded, Olfactory, Gene Expression

Journal: bioRxiv

Article Title: CXCL10 drives female-specific tau pathology progression and defines sex-dependent vulnerability in tauopathy model mice

doi: 10.64898/2026.04.19.719088

Figure Lengend Snippet: a Illustration explaining the definitions of cells neighboring Cxcl10 + astrocytes and Cxcl10 - astrocytes. b Volcano plot of gene expression showing Log₂ fold change > |0.25| and p < 0.1 highlighted in red, and Log₂ fold change < |0.25| and p > 0.1 shown in blue. All other transcripts are indicated in gray. c GO pathway analysis focusing on biological processes (BP) and molecular functions (MF) performed to compare neighboring cells of Cxcl10 + astrocytes with neighboring cells of Cxcl10 - astrocytes in male Tau Tg mice. The top 20 significantly upregulated pathways in neighboring cells of Cxcl10 + astrocytes are shown in each panel. Number of mice used: male Tau Tg (n = 1). ( b – d ) DEGs were identified using the Wilcoxon test, and GO analysis was evaluated via over-representation analysis employing the hypergeometric test. Source data are provided in the Source Data file.

Article Snippet: B6.129S4- Cxcl10 tm1Adl /J ( Cxcl10 KO, RRID:IMSR_JAX:006087) mice and B6.129X1- Mapt tm1Hnd /J ( Mapt KO, RRID:IMSR_JAX:007251) mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques: Gene Expression

Journal: bioRxiv

Article Title: CXCL10 drives female-specific tau pathology progression and defines sex-dependent vulnerability in tauopathy model mice

doi: 10.64898/2026.04.19.719088

Figure Lengend Snippet: b Representative images of CD3 + T cells in the hippocampi of male ( a ) and female ( b ) Tau Tg and Tau Tg; Cxcl10 KO mice aged 6 to 11–12 months. Scale bar = 500 μm. c, d Quantification of hippocampal T cells in Tau Tg and Tau Tg; Cxcl10 KO mice aged 6 to 11–12 months ( c : male, d : female). Data are presented as violin plots (n = 15–24 sections from 5–8 mice per group). Statistical analysis was performed using a two-way repeated measures ANOVA followed by a Šídák’s multiple comparisons test ( c ) and a mixed-effects analysis followed by a Šídák’s multiple comparisons test ( d ). ** p < 0.01, **** p < 0.0001. Source data are provided in the Source Data file.

Article Snippet: B6.129S4- Cxcl10 tm1Adl /J ( Cxcl10 KO, RRID:IMSR_JAX:006087) mice and B6.129X1- Mapt tm1Hnd /J ( Mapt KO, RRID:IMSR_JAX:007251) mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA).

Techniques:

Journal: Cell Reports Medicine

Article Title: Histone methyl-transferase G9a inhibition boosts the efficacy of immune checkpoint inhibitors in experimental hepatocellular carcinoma

doi: 10.1016/j.xcrm.2026.102717

Figure Lengend Snippet: G9a inhibition potentiates the immunogenic effects of IFN-γ in HCC cells (A) Effect of CM272 and EZM8266 on IFN-γ-triggered CXCL10 production in murine (PM299L) and human (HuH7) HCC cells. Cells were treated with CM272 for 48 h or with CM272 for 24 h and then with IFN-γ (75 U/mL) for another 24 h or with IFN-γ alone for 24 h. PM299L were treated with 400 nM, and HuH7 received 1 μM of CM272 ( n = 3). For EZM8266, cells were pretreated for 48 h with EZM8266 (5 mM) and then with IFN-γ (75 U/mL) for another 24 h, as indicated. CXCL10 protein levels were measured by ELISA in cells’ conditioned media ( n = 3). (B) Effect of G9a inhibition with CM272 or EZM8266 on the expression of MHC class I complex protein (MHC-I) on the surface of PM299L cells. Cells were treated with IFN-γ and CM272 or EZM8266 as indicated in (A), and MHC-I levels were determined by FACS analysis ( n = 3). (C) ChIP analyses of H3K9me2 levels in the proximal promoter regions of CXCL10 and HLA-A genes in HuH7 cells treated with IFN-γ (75 U/mL) and CM272 (400 nM), as indicated in (A) ( n = 3). (D) Evaluation of the expression of transposable elements (TEs) and endogenous retroviral sequences (ERVs) by RNA-seq in NM53 murine HCC cells treated with IFN-γ, CM272, and their combination as indicated in (A). (E) Immunofluorescence analyses of dsRNA in PM299L HCC cells treated with CM272 (24 h) or EZM8266 (48 h). Representative images are shown. Scale bars, 10 μm. Right panel shows a control without primary antibody ( n = 3). Data are represented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. All the replicates represent biological replicates.

Article Snippet: CXCL9 and CXCL10 concentrations were measured in culture supernatants collected at the end of the incubation periods using commercial ELISA kits for mouse CXCL9 (DY492) and CXCL10 (DY466), both from R&D Systems, and an ELISA kit for human CXCL10 (550926) from BD Biosciences (Franklin Lanes, NJ, USA).

Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Expressing, Retroviral, RNA Sequencing, Immunofluorescence, Control