Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 2. C-X-C motif chemokine 10 (CXCL10), stanniocalcin-1, and placental growth factor mRNA expression is significantly increased by trophoblast conditioned media (CM). Messenger RNA expression in vascular spheroids was measured by quantitative real-time PCR after stimulation with trophoblast CM for 24 hours (n=4). Data are expressed as mRNA expression relative to an external calibrator. A, CXCL10. B, Stanniocalcin-1. C, Placental growth factor. All data are presented as mean±SEM. *P<0.05.

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Expressing, RNA Expression, Real-time Polymerase Chain Reaction

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 3. C-X-C motif chemokine 10 (CXCL10) protein is expressed by vascular spheroids stimulated with trophoblast con- ditioned media. A, Vascular spheroid lysates were examined by Western blot analysis for the presence of CXCL10. Tubulin was used as an internal loading control. The image shown is represen- tative of 3 independent experiments. Densitometric analysis of Western blots (n=3) with mean±SEM CXCL10/tubulin presented as a fold-change over control-treated spheroids. *P<0.05. Vascu- lar spheroids treated with control media (B) or trophoblast condi- tioned media (C) were cryosectioned and sections were examined by immunostaining for the presence of CXCL10. Sections were taken through approximately the center of the spheroid. Negative control incubated with rabbit IgG in place of primary antibody is inset. Scale bar,100 µm.

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Western Blot, Control, Immunostaining, Negative Control, Incubation

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 4. C-X-C motif chemokine 10 (CXCL10) protein is expressed by first trimester decidua and dissected spiral arteries stimulated with trophoblast conditioned media. CXCL10 (A and C) and α-smooth muscle actin protein (D) expression was examined by immunohis- tochemistry in serially sectioned first trimester decidua (n=5; representative image shown). CXCL10 and α-smooth muscle actin protein colocalized to the same cells (indicated by arrowheads). Trophoblasts (labeled with CK7, B) were present at this stage of remodeling. Negative control (inset) was incubated with nonimmune IgG in place of primary antibody. Scale bar represents 100 µm or 50 µm in zoom. (E) Expression of CXCL10 (green) and (F) VWF, an endothelial cell marker (red), was examined in a dissected spiral artery treated with extravillous trophoblast (EVT) conditioned media (G, merge). EC indicates endothelial cells; and VSM, vascular smooth muscle. Negative control (inset) was incubated with nonimmune IgG in place of primary antibody. Scale bar, 50 µm.

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Expressing, Labeling, Negative Control, Incubation, Marker

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 5. The role of IFN-γ in C-X-C motif chemokine 10 (CXCL10) expression. A, Recombinant IFN-γ induces CXCL10 expression in vascular spheroids. Control or rhIFN-γ was added to spheroids made of endothelial cell (EC) alone, vascular smooth muscle cell (VSMC) alone, or cocultured EC/VSMC. CXCL10 expression was measured by Western blot analysis (n=4). B, Neutralizing IFN-γ in extravillous trophoblast (EVT) conditioned media (CM) decreased vascular spheroid CXCL10 expression. Control media or EVT CM was incubated with IFN-γ–neutralizing antibody or corresponding IgG control and added to EC/VSMC spheroids. CXCL10 expression was determined by Western blot analysis. *P<0.05 (n=5).

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Expressing, Recombinant, Control, Western Blot, Incubation

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Trophoblast-Induced Changes in C-X-C Motif Chemokine 10 Expression Contribute to Vascular Smooth Muscle Cell Dedifferentiation During Spiral Artery Remodeling

doi: 10.1161/atvbaha.112.300354

Figure Lengend Snippet: Figure 6. The effects of extravillous tro- phoblast (EVT) conditioned media (CM) and C-X-C motif chemokine 10 (CXCL10) on vascular smooth muscle cells (VSMCs). VSMCs were incubated for 72 hours in media containing 0.5% FCS, then a further 72 hours with indicated concentrations of EVT CM and recombi- nant human CXCL10 (n=at least 5 inde- pendent experiments). Expression of (A) α-smooth muscle actin and (B) calponin was examined by Western blot analysis. Time-lapse microscopy was used to analyze the effects of rhCXCL10 (C) and EVT CM (D) on VSMC motility during a 24-hour incubation. Data are displayed as the mean±SEM of a minimum of 3 pooled experiments. *P<0.05; **P<0.01.

Article Snippet: CXCL10 recombinant protein was purchased from R & D Systems (Abingdon, UK).

Techniques: Incubation, Expressing, Western Blot, Time-lapse Microscopy

Journal: Cellular and molecular biology (Noisy-le-Grand, France)

Article Title: Homocysteine modulates CXCL10/CXCR3 axis activity to induce endothelial dysfunction.

doi: 10.14715/cmb/2024.70.2.28

Figure Lengend Snippet: Fig. 3. (A, B) qRT-PCR and Western blot analysis of CXCL10 and CXCR3 expression in HAECs treated with different concentrations of Hcy. (C) IHC analysis of the aorta in HHcy mice showing CXCL10 and CXCR3 expression. (D) qRT-PCR analysis of CXCL10 and CXCR3 expression in arterial endothelial cells.

Article Snippet: Additionally, HAECs were exposed to specific agents including Anti-CXCL10 antibodies (701225, Invitrogen, USA), Anti-CXCR3 antibodies (ab71864, Abcam, UK), IgG control antibodies (31154, Thermo Fisher, USA), NBI-74330 (a CXCR3 inhibitor, 4528, Tocris Bioscience, UK), and CXCL10 agonist (266-IP, R&D Systems, USA).

Techniques: Quantitative RT-PCR, Western Blot, Expressing

Journal: Gut microbes

Article Title: Lacticaseibacillus paracasei sh2020 induced antitumor immunity and synergized with anti-programmed cell death 1 to reduce tumor burden in mice.

doi: 10.1080/19490976.2022.2046246

Figure Lengend Snippet: Figure 7. L. paracasei sh2020 promoted the expression and secretion of CXCL10 in vivo and in vitro. (a) The expression of CXCL9, CXCL10 and CXCL11 in tumor tissues from control and L. paracasei sh2020-treated mice was detected by qRT-PCR. (b-c) Representative images (b) and quantification (c) of IHC staining of CXCL10 in the tumor tissues from control and L. paracasei sh2020-treated tumors. (d) Tumor growth in each group. (e) The levels of CXCL10 in the conditioned medium. (f-g) Tumor growth in the tumor-bearing mice with intratumoral injection of L. paracasei sh2020 (n = 5–6). (h-i) Representative images (h) and quantification (i) of IHC staining of CXCL10 and CD8 in each group (n = 4–5). (j) The serum levels of CXCL10 were examined by ELISA. ns, no significant difference, *P < .05, **P < .01, ***P < .001.

Article Snippet: Depletion of T cell subsets and CXCL10 in vivo In the in vivo T cell subsets depletion study, InVivoMAb anti-mouse CD4 (YTS191, BE0119, BioXCell), CD8α (YTS169.4, BE0117 BioXcell) antibodies were intraperitoneally injected at a dose of 200 μg per mouse.

Techniques: Expressing, In Vivo, In Vitro, Control, Quantitative RT-PCR, Immunohistochemistry, Injection, Enzyme-linked Immunosorbent Assay

Journal: Gut microbes

Article Title: Lacticaseibacillus paracasei sh2020 induced antitumor immunity and synergized with anti-programmed cell death 1 to reduce tumor burden in mice.

doi: 10.1080/19490976.2022.2046246

Figure Lengend Snippet: Figure 8. CXCL10 controlled CD8+ T cell migration and the effect of L. paracasei sh2020 in vivo. (a-b) Representative images of IHC staining of CD8 and CXCL10 (a), and quantification (b) for the control and L. paracasei sh2020-treated tumors (n = 6–7). (c) IHC analysis of CD8 in tumors, which were divided into two groups according to CXCL10 high and low expression. (d) Experimental design: C57BL/6 mice were implanted subcutaneously with 5.0 × 105 MC38 cells and was treated with control vehicle or anti-CXCL10 antibody by intraperitoneal injection, every 3 days starting on D3, in total three times. The mice were given L. paracasei sh2020 with a dose of 1.0 × 109 CFU by gavage starting from D0 to D13. (e) Tumor growth in tumor-bearing mice in d. (f) Quantification of IHC staining of CXCL10 and CD8 in the tumors after neutralizing CXCL10 in vivo (n = 4–5). ns, no significant difference, *P < .05, **P < .01, ***P < .001.

Article Snippet: Depletion of T cell subsets and CXCL10 in vivo In the in vivo T cell subsets depletion study, InVivoMAb anti-mouse CD4 (YTS191, BE0119, BioXCell), CD8α (YTS169.4, BE0117 BioXcell) antibodies were intraperitoneally injected at a dose of 200 μg per mouse.

Techniques: Migration, In Vivo, Immunohistochemistry, Control, Expressing, Injection

Journal: Clinical Cancer Research

Article Title: XMT-2056, a HER2-Directed STING Agonist Antibody–Drug Conjugate, Induces Innate Antitumor Immune Responses by Acting on Cancer Cells and Tumor-Resident Immune Cells

doi: 10.1158/1078-0432.CCR-24-2449

Figure Lengend Snippet: XMT-2056 elicits potent antigen-dependent and Fc-R–mediated activation of monocytes in cocultures. A, Schematic of the XMT-2056 mechanism, which includes HER2-dependent ADC uptake into tumor cells and Fcγ-R–expressing myeloid cells. B, Chemical structure of XMT-2056. C, Cytokine induction as measured by a multiplex Luminex assay from supernatants of fresh human WBCs treated for 6 (IFN-β) or 24 (CXCL10, IL-6, and TNF-a) hours. Bars represent mean value of n = 2 data points shown as symbols. D, Competition with trastuzumab by biolayer interferometry (Octet): trastuzumab was loaded onto the sensor chip, and HER2 ECD or HT19 antibody associations are indicated by blue arrows. Additional binding by HT19 indicates noncompetitive binding. E, Competition with trastuzumab by cell-based flow cytometry. HT19 hIgG1 or mIgG2a formats detected with either Alexa Fluor anti-hIgG1 (h647) or Alexa Fluor anti-mIgG2a (m647) secondary antibodies in the presence or absence of trastuzumab–mIgG2a. Each point represents the mean and SD ( n = 3). F, Binding of XMT-2056 or HT19 antibody to HCC1954 cells showing fluorescence intensities measured by flow cytometry. Each point represents the mean and SD ( n = 2). G, CXCL10 cytokine induction in HCC1954 monocultures treated for 24 hours with XMT-2056, nonbinding control ADC, HT19, or the free payload. Each point represents the mean and SD ( n = 2). H, Binding of XMT-2056 or Fc-mutant XMT-2056, HT19, and nonbinding control ADC to SKOV3 cells showing fluorescence intensities measured by flow cytometry. Each point represents the mean and SD ( n = 3). I and J, IRF3 reporter activity of THP1 cells in coculture with SKOV3 cells ( I ) or cultured on recombinant HER2 antigen-coated plates ( J ) treated for 24 hours with XMT-2056 or Fc-mutant XMT-2056, nonbinding control ADC, or STING agonist payload. Each point represents the mean and SD ( n = 3). When noted, EC 50 and B max values represent mean of two independent experiments. gMFI, geometric mean fluorescence intensity; RLU, relative light units. ( A, Created in BioRender. Cetinbas, N. [2025], https://BioRender.com/s16c825 ; G, Created in BioRender. Cetinbas, N. [2025], https://BioRender.com/s16c825 ; I and J, Created in BioRender. Cetinbas, N. [2025], https://BioRender.com/s16c825 .)

Article Snippet: Culture supernatants were collected, and CXCL10 levels were measured by ELISA using Mouse CXCL10/IP-10 DuoSet ELISA Kit (R&D Systems, Cat. DY466) following the manufacturer’s recommended procedures.

Techniques: Activation Assay, Expressing, Multiplex Assay, Luminex, Binding Assay, Flow Cytometry, Fluorescence, Control, Mutagenesis, Activity Assay, Cell Culture, Recombinant