Journal: Signal Transduction and Targeted Therapy

Article Title: Ultra-high dose rate radiotherapy overcomes radioresistance in head and neck squamous cell carcinoma

doi: 10.1038/s41392-025-02184-0

Figure Lengend Snippet: The paracrine positive feedback loop between CD8 + T cells and macrophages induced by UHDR-RT. Venn diagram showing the intersecting genes/proteins obtained from transcriptome sequencing and proteomics analysis ( a ). IFN-γ and CXCL9 IF and IHC validation results ( b – d ). IF confirmed the colocalization of M1 macrophages with CXCL9 and of CD8 + T cells with IFN-γ ( e ). ELISAs verified the significant upregulation of CXCL9 and IFN-γ in the supernatants of radiated BMDMs and SPTCs, respectively ( f – h ). Mechanism diagram of UHDR-RT overcoming radioresistance. ( i ). Ctrl control; Conv-RT conventional radiotherapy; UHDR-RT ultra-high dose rate radiotherapy; BMDM bone marrow-derived macrophage; SPTC spleen-derived T cell; IHC immunohistochemistry; IF immunofluorescence; DSB double-strand break; CRT calreticulin; TiME tumor immune microenvironment; DC dendritic cell. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. The data are presented as the median and interquartile range ( c , d and g , h ). Comparisons were performed using one-way ANOVA with Tukey’s test for multiple comparisons. This figure was completed on the Figdraw platform ( https://www.figdraw.com/ )

Article Snippet: Primary antibodies included anti-mouse CXCL9 (#22355-1-AP, Proteintech, USA), CXCL10 (#10937-1-AP, Proteintech, USA), CXCL11 (#10707-1-AP, Proteintech, USA), CD8 (#GB15068, Servicebio, China), CD86 (#GB115630, Servicebio, China), and CD206 (#GB113497, Servicebio, China).

Techniques: Sequencing, Biomarker Discovery, Control, Derivative Assay, Immunohistochemistry, Immunofluorescence

Journal: Molecular Pain

Article Title: Spinal CXCL9 and CXCL11 are not involved in neuropathic pain despite an upregulation in the spinal cord following spinal nerve injury

doi: 10.1177/1744806918777401

Figure Lengend Snippet: Primer sets used in qPCR.

Article Snippet: The sections were first blocked with 5% donkey serum for 2 h at room temperature, then incubated overnight at 4°C with the following primary antibodies: CXCL9 (Rabbit, 1:500, Bio-Rad), CXCL11 (Rabbit, 1:500, Bio-Rad), glial fibrillary acidic protein (GFAP; mouse, 1:5000, Millipore), NeuN (mouse, 1:1000, Millipore), and CD11b (mouse, 1:100, Serotec).

Techniques: Sequencing

Journal: Molecular Pain

Article Title: Spinal CXCL9 and CXCL11 are not involved in neuropathic pain despite an upregulation in the spinal cord following spinal nerve injury

doi: 10.1177/1744806918777401

Figure Lengend Snippet: The CXCL9 expression is increased in spinal astrocytes after SNL. (a) Time course of Cxcl9 mRNA expression in the ipsilateral dorsal horn in naive, sham-operated, and SNL mice. Cxcl9 expression was significantly increased at 3, 10, and 21 days in SNL mice. *p < 0.05, **p < 0.01, compared with sham-operated mice. Student's t test. n = 5 mice/group. (b) ELISA shows the increase of CXCL9 protein in the spinal cord 10 days after SNL. *p < 0.05, compared with sham-operated mice. Student’s t test, n = 5 mice/group. (c to d) Representative images of CXCL9 immunofluorescence in the spinal cord from naïve and SNL mice, receptively. CXCL9 was constitutively expressed in naive mice (c), but significantly increased in the ipsilateral dorsal horn 10 days after SNL mice (d). (e) CXCL9-IR was not shown after absorption with CXCL9 peptide. (f to h) Double staining shows the cellular distribution of CXCL9 in the spinal dorsal horn. CXCL9 was sparely colocalized with NeuN (f) or CD11b (g), but highly colocalized with GFAP (h) in the spinal cord 10 days after SNL.

Article Snippet: The sections were first blocked with 5% donkey serum for 2 h at room temperature, then incubated overnight at 4°C with the following primary antibodies: CXCL9 (Rabbit, 1:500, Bio-Rad), CXCL11 (Rabbit, 1:500, Bio-Rad), glial fibrillary acidic protein (GFAP; mouse, 1:5000, Millipore), NeuN (mouse, 1:1000, Millipore), and CD11b (mouse, 1:100, Serotec).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Double Staining

Journal: Molecular Pain

Article Title: Spinal CXCL9 and CXCL11 are not involved in neuropathic pain despite an upregulation in the spinal cord following spinal nerve injury

doi: 10.1177/1744806918777401

Figure Lengend Snippet: Inhibition of CXCL9 or CXCL11 does not alleviate SNL-induced neuropathic pain. (a to c) The shRNA targeting Cxcl9 , Cxcl11 , or Cxcl10 reduced the mRNA expression of Cxcl9 (a), Cxcl11 (b), and Cxcl10 (c) in HEK293 cells. (d to e) Intraspinal injection of LV- Cxcl9 shRNA or LV- Cxcl11 shRNA three days after SNL did not change the paw withdrawal latency (d) or threshold (e) at days 10 and 14. p > 0.05, two-way RM ANOVA. n = 6–7 mice/group. (f to g) Intraspinal injection of LV- Cxcl10 shRNA three days after SNL significantly increased the paw withdrawal latency (f) and threshold (g) at days 10 and 14. ** p < 0.01, *** p <0.001, two-way RM ANOVA followed by Bonferroni’s test, n = 5–6 mice/group.

Article Snippet: The sections were first blocked with 5% donkey serum for 2 h at room temperature, then incubated overnight at 4°C with the following primary antibodies: CXCL9 (Rabbit, 1:500, Bio-Rad), CXCL11 (Rabbit, 1:500, Bio-Rad), glial fibrillary acidic protein (GFAP; mouse, 1:5000, Millipore), NeuN (mouse, 1:1000, Millipore), and CD11b (mouse, 1:100, Serotec).

Techniques: Inhibition, shRNA, Expressing, Injection

Journal: Molecular Pain

Article Title: Spinal CXCL9 and CXCL11 are not involved in neuropathic pain despite an upregulation in the spinal cord following spinal nerve injury

doi: 10.1177/1744806918777401

Figure Lengend Snippet: CXCL9 and CXCL11 do not induce pain hypersensitivity after intrathecal injection. (a, b) Intrathecal injection of CXCL9 and CXCL11 (100 ng) did not induce heat hyperalgesia (a) or mechanical allodynia (b) in naive mice. p > 0.05, two-way RM ANOVA. n = 6 mice/group. (c, d) Intrathecal injection of CXCL10 (100 ng) induced heat hyperalgesia (C) at 1, 3, and 6 h and mechanical allodynia (d) at 1 h and 3 h in naive mice. *** p < 0.001, two-way RM ANOVA followed by Bonferroni’s test, n = 6–7 mice/group. (e) pERK expression in the spinal cord did not change 1 h after intrathecal injection of CXCL9 or CXCL11 in naive mice. p > 0.05, Student's t test, n = 4 mice/group.

Article Snippet: The sections were first blocked with 5% donkey serum for 2 h at room temperature, then incubated overnight at 4°C with the following primary antibodies: CXCL9 (Rabbit, 1:500, Bio-Rad), CXCL11 (Rabbit, 1:500, Bio-Rad), glial fibrillary acidic protein (GFAP; mouse, 1:5000, Millipore), NeuN (mouse, 1:1000, Millipore), and CD11b (mouse, 1:100, Serotec).

Techniques: Injection, Expressing

Journal: Molecular Pain

Article Title: Spinal CXCL9 and CXCL11 are not involved in neuropathic pain despite an upregulation in the spinal cord following spinal nerve injury

doi: 10.1177/1744806918777401

Figure Lengend Snippet: CXCL9 and CXCL11 enhance excitatory synaptic transmission in lamina II neurons. (a) Whole-cell patch clamp recording of sEPSCs shows an increase in the frequency of sEPSCs after perfusion of CXCL9 (100 ng/ml, 2 min). However, the amplitude was not changed after perfusion of CXCL9. a1 and a2 are enlarged traces before and after CXCL9 treatment, respectively. ***p < 0.001 versus pretreatment baseline, Student’s t test, n = 5–6 mice/group. (b) sEPSCs frequency not amplitude is increased after perfusion of CXCL11 (100 ng/ml, 2 min). b1 and b2 are enlarged traces before and after CXCL11 treatment, respectively. **p < 0.01 versus pretreatment baseline, Student’s t test, n = 5–6 mice/group. (c) Miniature EPSCs (mEPSCs) were recorded in the present of TTX (500 nM). The mEPSCs frequency, not the amplitude is increased after perfusion of CXCL9 (100 ng/ml, 2 min). c1 and c2 are enlarged traces before and after CXCL9 treatment, respectively. *p < 0.05 versus pretreatment baseline, Student’s t test, n = 3 mice/group. (d) mEPSCs frequency, not the amplitude is increased after incubation of CXCL11. d1 and d2 are enlarged traces before and after CXCL11 treatment, respectively. * p < 0.05 versus pretreatment baseline, Student's t test, n = 3 mice/group.

Article Snippet: The sections were first blocked with 5% donkey serum for 2 h at room temperature, then incubated overnight at 4°C with the following primary antibodies: CXCL9 (Rabbit, 1:500, Bio-Rad), CXCL11 (Rabbit, 1:500, Bio-Rad), glial fibrillary acidic protein (GFAP; mouse, 1:5000, Millipore), NeuN (mouse, 1:1000, Millipore), and CD11b (mouse, 1:100, Serotec).

Techniques: Transmission Assay, Patch Clamp, Incubation

Journal: Molecular Pain

Article Title: Spinal CXCL9 and CXCL11 are not involved in neuropathic pain despite an upregulation in the spinal cord following spinal nerve injury

doi: 10.1177/1744806918777401

Figure Lengend Snippet: CXCL9 and CXCL11 enhance inhibitory synaptic transmission in lamina II neurons. (a) Patch clamp recording shows an increase in the frequency but not the amplitude of the sIPSC after perfusion of CXCL9 (100 ng/ml, 2 min). a1 and a2 are enlarged traces before and after CXCL9 treatment. *p < 0.05, Student’s t test, n = 3–4 mice/group. (b) The frequency, not the amplitude of the sIPSCs was increased during perfusion of CXCL11 (100 ng/ml, 2 min). b1 and b2 are enlarged traces before and after CXCL11 treatment. ***p < 0.001, Student’s t test, n = 3–4 mice/group. (c) Neither the frequency nor the amplitude of the sIPSCs was changed after perfusion of CXCL10 (100 ng/ml, 2 min). c1 and c2 are enlarged traces before and after CXCL10 treatment. p > 0.05, Student’s t test, n = 3–4 mice/group.

Article Snippet: The sections were first blocked with 5% donkey serum for 2 h at room temperature, then incubated overnight at 4°C with the following primary antibodies: CXCL9 (Rabbit, 1:500, Bio-Rad), CXCL11 (Rabbit, 1:500, Bio-Rad), glial fibrillary acidic protein (GFAP; mouse, 1:5000, Millipore), NeuN (mouse, 1:1000, Millipore), and CD11b (mouse, 1:100, Serotec).

Techniques: Transmission Assay, Patch Clamp

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: CXCL9 influences the tumor immune microenvironment by stimulating JAK/STAT pathway in triple-negative breast cancer

doi: 10.1007/s00262-022-03343-w

Figure Lengend Snippet: ROC curves of the a ADAMTS5, b TACC3, c HOXA4, d ABCA5, and e CXCL9 in the training dataset

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, the membranes were incubated overnight with the primary antibodies CXCL9 (1:500, Proteintech, China), JAK1 (1:1000), and STAT2 (1:1000) at 4 °C.

Techniques:

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: CXCL9 influences the tumor immune microenvironment by stimulating JAK/STAT pathway in triple-negative breast cancer

doi: 10.1007/s00262-022-03343-w

Figure Lengend Snippet: Overlapping genes and enrichment analysis. a Red dots indicate high expression of the DEGs in the GSE157284 volcano plot; b a Venn diagram showing overlapping genes between the PD-L1 expression status-DEGs, WGCNA-yellow genes, LASSO, and SVM; c and d expression of CXCL9 in TNBC para-cancerous and TNBC cancerous tissues determined with immunohistochemistry staining; e log-rank test with Kaplan–Meier survival curves was performed to compare overall survival (OS) between breast cancer patients with high and low CXCL9 expression; f log-rank test with Kaplan–Meier survival curves was performed to compare overall survival (OS) between TNBC patients with high and low CXCL9 expression groups

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, the membranes were incubated overnight with the primary antibodies CXCL9 (1:500, Proteintech, China), JAK1 (1:1000), and STAT2 (1:1000) at 4 °C.

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: CXCL9 influences the tumor immune microenvironment by stimulating JAK/STAT pathway in triple-negative breast cancer

doi: 10.1007/s00262-022-03343-w

Figure Lengend Snippet: Identification of different cell clusters in TNBC by single-cell sequencing analysis. a Distribution of immune cell-type fractions in the PD‐L1 expression status in TNBC; b the proportions of 22 TIICs in the PD-L1-positive (red) and PD-L1-negative (blue) TNBC; c correlation between the expression of CXCL9 and the infiltration of immune cells from TNBC; d comparison of batch effects between the groups; e PCA data showing the 50 PCs with a P value < 0.05; f the 16 cell clusters classified using the UMAP algorithm; g the 16 cell clusters annotated into major types using SingleR and CellMarker; h and i UMAP and violin plots showing high expression of CXCL9 gene in M1 macrophages

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, the membranes were incubated overnight with the primary antibodies CXCL9 (1:500, Proteintech, China), JAK1 (1:1000), and STAT2 (1:1000) at 4 °C.

Techniques: Sequencing, Expressing, Comparison

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: CXCL9 influences the tumor immune microenvironment by stimulating JAK/STAT pathway in triple-negative breast cancer

doi: 10.1007/s00262-022-03343-w

Figure Lengend Snippet: Cell cluster trajectory and CellChat analysis. a Heatmap of different cell clusters interaction correlation; b the cellular interaction weights of interactions between M1 macrophages and M2 macrophages; c the differentiation pseudotime trajectory of M1 macrophages and M2 macrophages; d pseudotime was colored in a gradient from dark blue to light blue; e the expression intensity of CXCL9 along the pseudotime axis; f the MHC-II signaling pathway network was significantly detected between the interactions of M1 macrophage and M2 macrophage; g differences in pathway activities scored by GSVA between all types of cell types; h and i GSEA analysis revealed a significant enrichment of MHC-II (hsa 04,514) and JAK/STAT signaling pathways. NES, normalized enrichment score; j CXCL9 was upregulated in the pENTER-CXCL9 group. JAK1 and STAT2 expressions were higher in the pENTER-CXCL9 group than in the control group

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, the membranes were incubated overnight with the primary antibodies CXCL9 (1:500, Proteintech, China), JAK1 (1:1000), and STAT2 (1:1000) at 4 °C.

Techniques: Expressing, Protein-Protein interactions, Control