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: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Phenotypic polarization of activated astrocytes: the critical role of lipocalin-2 in the classical inflammatory activation of astrocytes.

doi: 10.4049/jimmunol.1301637

Figure Lengend Snippet: FIGURE 1. Identification of phenotypic polarization in primary astrocyte cultures based on gene-expression profiles. Primary astrocyte cultures were treated with LPS (100 ng/ml) plus IFN-g (50 U/ml), IL-4 (10 ng/ml), ICs (OVA [75 mg/ml] plus anti- OVA Ab [150 mg/ml]) plus LPS (100 ng/ ml), or IL-10 (10 ng/ml) for 8 h, and total RNAs were isolated. (A) The mRNA levels of phenotypic markers (Il-1b, Inos, Tnf-a, Cxcl10, Il-12, Il-23, Il-10, Arg1, Mrc1, Il- 1ra, Fizz1, and Ym1) were determined by real-time RT-PCR. (B) Polarized classical versus alternative activation was also as- sessed by the ratio of expressed mRNA markers (Il-1b/Il-1ra, Inos/Arg1, Tnf-a /Il- 10). Gapdh was used as an internal control. Results are mean 6 SD (n = 3). *p , 0.05, versus untreated control.

Article Snippet: Recombinant mouse TNF-a and CXCL10 protein (R&D Systems) were used as standards.

Techniques: Gene Expression, Isolation, Quantitative RT-PCR, Activation Assay, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Phenotypic polarization of activated astrocytes: the critical role of lipocalin-2 in the classical inflammatory activation of astrocytes.

doi: 10.4049/jimmunol.1301637

Figure Lengend Snippet: FIGURE 2. Assessment of NO pro- duction, TNF-a/CXCL10 secretion, ex- pression of ARG1/MRC1 proteins, and GFAP levels following exposure to ei- ther classical or alternative activation stimuli. Primary astrocyte cultures were treated with LPS (100 ng/ml) plus IFN- g (50 U/ml), IL-4 (10 ng/ml), IC (OVA [75 mg/ml] plus OVA Ab [150 mg/ml]) plus LPS (100 ng/ml), or IL-10 (10 ng/ ml) for 24 h. (A) Concentrations of ni- trite, TNF-a protein, and CXCL10 pro- tein in culture media were measured using Griess reagent or specific ELISA. (B) ARG1 and MRC1 protein expres- sion was detected by flow cytometry using Abs specific for ARG1 or MRC1. Mean fluorescence intensity (MFI) val- ues are also shown. Negative control is the measurement without specific Ab or with a control Ab. GFAP mRNA or protein levels were evaluated by RT- PCR (C) or Western blotting (D), re- spectively. Gapdh or a-tubulin was used as an internal control. Results are mean 6 SD (n = 3). *p , 0.05, versus un- treated control.

Article Snippet: Recombinant mouse TNF-a and CXCL10 protein (R&D Systems) were used as standards.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Cytometry, Negative Control, Control, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Physiological Reports

Article Title: CXCL10 is a novel anti‐angiogenic factor downstream of p53 in cardiomyocytes

doi: 10.14814/phy2.15304

Figure Lengend Snippet: p53 induced CXCL10 expression in cardiomyocytes. DNA microarray was performed using p53‐overexpressing NRCMs. (a) Scatter plot shows the genes colored blue with >2‐fold upregulate in p53‐overexpressing NRCMs compared with β‐gal. (b) Heat map shows genes colored red with >2‐fold change and green with <0.5‐fold change between β‐gal and p53. (c) Gene ontology analysis. (d, e) The expression of (d) CXCL10 and (e) p21 transcripts was measured by qPCR ( n = 4). Experiments were performed three times with similar results. (f) The protein expression of CXCL10 in culture media of p53‐overexpressing NRCMs was measured by ELISA ( n = 3). * p < 0.05, ** p < 0.01. by Student's t ‐test.

Article Snippet: CXCL10 protein level in conditioned media, secreted from cardiomyocytes, was assessed by Rat IP‐10/CXCL10 ELISA Kit (Elabscience, #E‐EL‐R0546) according to the manufacturer's protocol.

Techniques: Expressing, Microarray, Enzyme-linked Immunosorbent Assay

Journal: Physiological Reports

Article Title: CXCL10 is a novel anti‐angiogenic factor downstream of p53 in cardiomyocytes

doi: 10.14814/phy2.15304

Figure Lengend Snippet: Doxorubicin induced the expression of CXCL10 in cardiomyocytes through p53 in cardiomyocytes. (a–h) NRCMs were stimulated with Doxo for indicated concentration and time. (i–l) NRCMs were transfected with siRNA for p53 (sip53) or control (siCon) for 48 h, followed by stimulation with Doxo for 24 h. The expression of p53 (a, e, i) and CXCL10 (b, f, j) mRNA was measured by qPCR. ( n = 3). (c, g, k) The expression of p53 protein was analyzed by immunoblotting. Representative data are shown. (d, h, l) The band intensities of p53 were measured ( n = 3). Experiments were performed three times with similar results. (a–h). *p < 0.05, **p < 0.01 by one‐way ANOVA followed by Dunnett test. (i–l) * p < 0.05, ** p < 0.01 by one‐way ANOVA followed by Tukey–Kramer test.

Article Snippet: CXCL10 protein level in conditioned media, secreted from cardiomyocytes, was assessed by Rat IP‐10/CXCL10 ELISA Kit (Elabscience, #E‐EL‐R0546) according to the manufacturer's protocol.

Techniques: Expressing, Concentration Assay, Transfection, Control, Western Blot

Journal: Physiological Reports

Article Title: CXCL10 is a novel anti‐angiogenic factor downstream of p53 in cardiomyocytes

doi: 10.14814/phy2.15304

Figure Lengend Snippet: Hypoxia synergistically elevated the expression of CXCL10. p53‐overexpresisng NRCMs were treated with CoCl2 for 24 h. (a) The expression of p53 and GAPDH protein was analyzed by immunoblotting. Representative data were shown. The expression of (b) VEGF and (c) CXCL10 transcripts was measured by qPCR ( n = 4). Experiments were performed three times with similar results. * p < 0.05, ** p < 0.01 by one‐way ANOVA followed by Tukey–Kramer test.

Article Snippet: CXCL10 protein level in conditioned media, secreted from cardiomyocytes, was assessed by Rat IP‐10/CXCL10 ELISA Kit (Elabscience, #E‐EL‐R0546) according to the manufacturer's protocol.

Techniques: Expressing, Western Blot

Journal: Physiological Reports

Article Title: CXCL10 is a novel anti‐angiogenic factor downstream of p53 in cardiomyocytes

doi: 10.14814/phy2.15304

Figure Lengend Snippet: p53‐induced CXCL10, secreted from cardiomyocytes, inhibited the tube formation of endothelial cells. Culture media were collected from p53‐ or β‐gal‐overexpressing NRCMs. RAOECs were cultured with the conditioned media in the presence of AMG487, an inhibitor of CXCR3. Angiogenesis assay was performed. (a) Representative images were shown. (b) The number of tube formation was measured ( n = 6). * p < 0.05, ** p < 0.01 by one‐way ANOVA followed by Tukey–Kramer test.

Article Snippet: CXCL10 protein level in conditioned media, secreted from cardiomyocytes, was assessed by Rat IP‐10/CXCL10 ELISA Kit (Elabscience, #E‐EL‐R0546) according to the manufacturer's protocol.

Techniques: Cell Culture, Angiogenesis Assay

Journal: Journal of nanobiotechnology

Article Title: Unveiling the improved targeting migration of mesenchymal stem cells with CXC chemokine receptor 3-modification using intravital NIR-II photoacoustic imaging.

doi: 10.1186/s12951-022-01513-7

Figure Lengend Snippet: Fig. 4 Upregulation of Cxcl10 in inflamed ears of CHS mice. a Representative photographs of mouse ears challenged with vehicle (Control) or DNFB (Inflamed). Photos were taken 24 h after challenging. b Thickness of mice ears challenged with vehicle (Control) or DNFB (Inflamed) before (Pre), 24 h and 72 h after challenging (n = 3–4 per group). c mRNA expression of Cxcl10 transcript in mouse ears challenged with vehicle (Control) or DNFB (Inflamed) (n = 3 per group). ***P < 0.001; ns, no significance; error bars, SEM

Article Snippet: Rabbit polyclonal antibody against Cxcr3 (NB100-56404), recombinant Cxcl10 protein (466- CR) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Control, Expressing

Journal: Journal of nanobiotechnology

Article Title: Unveiling the improved targeting migration of mesenchymal stem cells with CXC chemokine receptor 3-modification using intravital NIR-II photoacoustic imaging.

doi: 10.1186/s12951-022-01513-7

Figure Lengend Snippet: Fig. 5 Overexpression of Cxcr3 in MSCs. a Representative bright field (BF) and green fluorescence images of MSCeGFP and MSCCxcr3. MSCs were transduced with lentivirus to express eGFP alone (MSCeGFP) or in combination with Cxcr3 (MSCCxcr3). b Analysis of Cxcr3 mRNA expression in MSCeGFP and MSCCxcr3.c Western blot of Cxcr3 in total cell lysates of MSCeGFP and MSCCxcr3. CypB, cyclophilin B, used as housekeeping gene. d Cxcr3 overexpression promoted the in vitro migration of MSCs toward Cxcl10. Representative images and quantification analysis of transwell migration assay were shown. e Quantification analysis of transwell migration assay of MSCCxcr3 after incubating with vehicle (PBS) or TAT-CPNPs (NPs). Scale bars, 100 µm. *P < 0.05, ***P < 0.001; error bars, SEM

Article Snippet: Rabbit polyclonal antibody against Cxcr3 (NB100-56404), recombinant Cxcl10 protein (466- CR) were purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Over Expression, Fluorescence, Transduction, Expressing, Western Blot, In Vitro, Migration, Transwell Migration Assay