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: Journal of Advanced Research

Article Title: The pro-cancer and immunosuppressive activity of macrophage-transformed cancer-associated fibroblasts in oral squamous cell carcinoma

doi: 10.1016/j.jare.2025.07.027

Figure Lengend Snippet: MMTs recruit a large number of MDSCs through CXCL10 pathway. A Significantly upregulated genes in MMTs versus non-MMT fibroblasts from OSCC database (GSA-Human: HRA007439). B Differences in CXCL10 expression between non-MMT fibroblasts and MMTs were demonstrated in HNSCC (figure S1). C CXCL10 expression and release were detected by PCR and ELISA. D-H Exogenous CXCL10 was added to BMDMs, while the CXCL10 inhibitor AMG487 was administered to MMTs, followed by co-culturing with MDSCs. The migration of MDSCs was evaluated using transwell chambers and crystal violet staining ( D ). Flow cytometry was used to detect cell apoptosis ( E ), macrophage markers (F4/80 + CD11b + ) ( F ), as well as CFSE for assessing T cell proliferation inhibition ( G ). PCR and ELISA were employed to measure the expression of immunosuppressive factors IL10 and iNOS ( H ). I, J Tumor tissue photographs ( I ) and the tumor growth curve were taken and subjected to statistical analysis regarding volume and weight ( J ). K-M Flow cytometry was used to detect the proportion of MDSCs in the spleen ( K ) and tumors ( L ) of each group, while immunofluorescence staining ( M ) revealed the densest field of MDSCs within the tumor tissues across all groups. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs experimental group. “ns” represents no statistical significance. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: According to the manufacturer's instructions, the cytokine concentrations of TGFβ1, iNOS, IL-10, and CXCL10 in the supernatant were determined using ELISA kits (Boster, California, USA).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Migration, Staining, Flow Cytometry, Inhibition, Immunofluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: Activation of the STING‐IRF3 pathway involved in psoriasis with diabetes mellitus

doi: 10.1111/jcmm.17236

Figure Lengend Snippet: STING‐IRF3 pathway involvement in cellular model of diabetes and psoriasis. (A, B) Protein levels of STING, p‐TBK1/TBK1, p‐IRF3/IRF3, CXCL10 and IFN‐β were measured by Western blotting. (C) The expression level of CXCL10 in HaCaT cells was measured by ELISA. Data are represented as means ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001

Article Snippet: The mouse serum and HaCaT cell culture media of each group were further analysed to determine the concentrations of interferon gamma‐induced protein 10 (CXCL10) and IFN‐β using ELISA kits, according to the manufacturer's instructions (Elabscience) (Mouse IFN‐β ELISA Kit: E‐EL‐M0033c, Mouse CXCL10 ELISA Kit: E‐EL‐M0021c, Human CXCL10 ELISA Kit: E‐EL‐H0050c).

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Activation of the STING‐IRF3 pathway involved in psoriasis with diabetes mellitus

doi: 10.1111/jcmm.17236

Figure Lengend Snippet: Evaluation of STING‐IRF3 pathway in psoriasis and T2DM animal model. Group A, NCD + Vaseline, Group B, NCD + IMQ, Group C, DM + Vaseline, Group D, DM + IMQ. (A) The protein levels of STING, p‐IRF3, IL‐17A and IL‐23 in the skin tissue were measured by immunohistochemistry (×200). (B, C) The protein levels of STING, p‐TBK1/TBK1, p‐IRF3/IRF3 and CXCL10, and inflammatory cytokines IFN‐β, NF‐κB p65, TNF‐α, pro‐IL‐1β, IL‐17A and IL‐23 in skin tissue were measured by Western blotting. (D) Serum CXCL10 and IFN‐β of mice were measured by ELISA. Data are represented as means ± SEM ( n = 6). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; Scale bars, 50 µm

Article Snippet: The mouse serum and HaCaT cell culture media of each group were further analysed to determine the concentrations of interferon gamma‐induced protein 10 (CXCL10) and IFN‐β using ELISA kits, according to the manufacturer's instructions (Elabscience) (Mouse IFN‐β ELISA Kit: E‐EL‐M0033c, Mouse CXCL10 ELISA Kit: E‐EL‐M0021c, Human CXCL10 ELISA Kit: E‐EL‐H0050c).

Techniques: Animal Model, Immunohistochemistry, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Activation of the STING‐IRF3 pathway involved in psoriasis with diabetes mellitus

doi: 10.1111/jcmm.17236

Figure Lengend Snippet: Inhibition of STING‐IRF3 pathway in diabetic mice with psoriasis. Group E, NCD + Vaseline, Group F, DM + IMQ, Group G, DM + IMQ + C‐176. (A) Experimental procedure for the STING inhibitor: C‐176. After 7 days of intervention with IMQ, (B) visual changes in the skin on the back of each group. (C) Comparison of PASI scores of back skin of mice in each group. (D) Histopathological changes were observed under light microscope after HE staining (×200). (E) The epidermal thickness and the number of inflammatory cells in the dermis were analysed by Image‐Pro Plus 6.0 software (×200). (F) Protein levels of STING, p‐IRF3, IL‐17A and IL‐23 in skin tissue were measured by immunohistochemistry (×200). (G) Protein levels of STING, p‐TBK1/TBK1, p‐IRF3/IRF3 and CXCL10, and inflammatory cytokines IFN‐β, NF‐κB p65, TNF‐α, pro‐IL‐1β, IL‐17A and IL‐23 in skin tissue were measured by Western blotting. (H) Total mRNA levels of STING and IFN‐β in skin tissue were measured by qRT‐PCR. (I) Serum CXCL10 and IFN‐β of mice were measured by ELISA. Data are represented as means ± SEM ( n = 6 mice in each group). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Scale bars, 50 µm

Article Snippet: The mouse serum and HaCaT cell culture media of each group were further analysed to determine the concentrations of interferon gamma‐induced protein 10 (CXCL10) and IFN‐β using ELISA kits, according to the manufacturer's instructions (Elabscience) (Mouse IFN‐β ELISA Kit: E‐EL‐M0033c, Mouse CXCL10 ELISA Kit: E‐EL‐M0021c, Human CXCL10 ELISA Kit: E‐EL‐H0050c).

Techniques: Inhibition, Comparison, Light Microscopy, Staining, Software, Immunohistochemistry, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Pathogen evasion of chemokine response through suppression of CXCL10

doi: 10.1101/557876

Figure Lengend Snippet: (A) Cytokine screening of LCLs exposed to L. major demonstrated suppression of CXCL10. Three lymphoblastoid cell lines (LCL), 7357, 18524, and 19203, were infected with L. major. Chemokines secreted into culture supernatants were analyzed by Luminex. Cytokines below the limit of detection were removed from the final analysis. Values are represented as log 2 of the fold change relative to uninfected LCLs. Type-1 associated cytokines are represented in grey. P value represents Dunnett’s post-hoc test compared to 1, after repeated measures one-way ANOVA. (B) CXCL10 produced by LPS stimulated THP-1 monocytes was suppressed by L. major. THP-1 monocytes were stimulated with LPS prior to L. major infection. CXCL10 mRNA was measured by qRT-PCR TaqMan assay using the ΔΔC t method with 18s as housekeeping gene, and CXCL10 protein was measured by ELISA. For mRNA (n=3) and ELISA (n=6), Fold Change is relative to unstimulated, uninfected THP-1s. P values calculated by Student’s t-test .

Article Snippet: The CXCL10 plasmid was obtained from Origene (NM_001565), and contains C-terminal Myc and Flag epitope tags.

Techniques: Infection, Luminex, Produced, Quantitative RT-PCR, TaqMan Assay, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Pathogen evasion of chemokine response through suppression of CXCL10

doi: 10.1101/557876

Figure Lengend Snippet: (A) Zinc chelation prevents CXCL10 suppression. Concentration of human recombinant CXCL10 was measured by ELISA after incubation for 12 hours with filtered conditioned media from L. major WT promastigote culture and addition of the zinc-chelator 1,10-phenanthroline (n=8). (B) gp63 is required for L. major CXCL10 suppression. Human recombinant CXCL10 concentrations were measured by ELISA after 12 hour incubation with conditioned media from L. major WT, Δ gp63 , or Δ gp63 + 1 (n=6). (C) GP63 expressed and secreted by HEK293Ts is sufficient for CXCL10 suppression. Human recombinant CXCL10 concentrations were measured by ELISA after 12 hour incubation with culture supernatant from HEK293Ts transfected with pCDNA3.1-gp63 WT or pCDNA3.1-gp63 E285A (n=7). P values calculated by one-way ANOVA with Tukey’s post-hoc test. Error bars represent standard error of the mean.

Article Snippet: The CXCL10 plasmid was obtained from Origene (NM_001565), and contains C-terminal Myc and Flag epitope tags.

Techniques: Concentration Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Incubation, Transfection

Journal: bioRxiv

Article Title: Pathogen evasion of chemokine response through suppression of CXCL10

doi: 10.1101/557876

Figure Lengend Snippet: (A) CXCL9/10/11 share significant homology at the amino-acid level. Multisequence alignment demonstrates that physical characteristics of amino acids are conserved across the CXCL10 family of chemokines. There are three putative GP63 cleavage sites (underlined) based on the consensus sequence of polar (P1), hydrophobic (P1’), basic (P2’) . (B) GP63 selectively cleaves chemokine ligands of the CXCR3 receptor. Conditioned media from L. major WT, Δgp63 , and Δgp63 + 1 was incubated with human recombinant chemokines for 12 hours and product detected by western blot. Cleavage is only detected for the CXCL9/10/11 family. (C) Cleavage by GP63 generates a smaller molecular weight protein. A time course of cleavage of human CXCL10 by heterologously expressed GP63 demonstrated an intermediate cleavage product, resolved by PAGE and Coomassie staining. (D) Cleavage by GP63 results in a change in CXCL10 molecular weight of 2.2kD. Capillary electrophoresis-Mass Spectrometry (CE-MS) determined the molecular weight of the uncleaved (CXCL10 Hi ) and cleaved (CXCL10 Lo ) bands as 8.8kD and 6.6kD respectively. (E) Comparative analysis by trypsin digest of cleaved and uncleaved CXCL10 reveals cleavage occurring between A81-I82. Liquid chromatography-mass spectrometry (LC-MS) following trypsin digest of CXCL10 Hi and CXCL10 Lo identified peptide ending at A81, exclusively in the CXCL10 Lo band, and a corresponding lack of peptide coverage from AA84-91. (F) Mutation of A81F significantly impairs GP63 cleavage of CXCL10. In the presence of GP63, CXCL10 A81F remains stable for up to 45 minutes whereas CXCL10 WT degradation is nearly complete by 15 minutes. Percentage of GP63 remaining at 15 minutes is plotted (n=3-4 per CXCL10 genotype). P value calculated by Student’s t-test . (G) The GP63 cleavage site is found on the C-terminal alpha-helix loop of CXCL10. Based on the NMR crystal structure of CXCL10 (Booth et al., 2002), the A81, I82, K83 (P1, P1’, P2’) GP63 cleavage motif maps to an exposed alpha-helical region.

Article Snippet: The CXCL10 plasmid was obtained from Origene (NM_001565), and contains C-terminal Myc and Flag epitope tags.

Techniques: Sequencing, Incubation, Recombinant, Western Blot, Molecular Weight, Staining, Electrophoresis, Mass Spectrometry, Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Mutagenesis

Journal: bioRxiv

Article Title: Pathogen evasion of chemokine response through suppression of CXCL10

doi: 10.1101/557876

Figure Lengend Snippet: (A) L. major promastigotes suppress CXCL10 through GP63 activity. THP-1 monocytes were differentiated using 100ng/mL of PMA prior to infection and CXCL10 concentration was assessed in the supernatant 24 hours post-infection by ELISA. (B) L. major amastigotes suppress CXCL10 through GP63 activity. At 24 hours post-infection, extracellular promastigotes were washed away from the differentiated THP-1 monocytes. At 48 hours post-infection the CXCL10 concentration was assessed in the supernatant by ELISA. For A-B, data represents four separate infections and was analyzed by one-way ANOVA with Tukey’s post-hoc test (C) L. major induces similar levels of CXCL10 mRNA, independent of GP63 genotype. At 48 hours post-infection, mRNA was extracted from PMA differentiated THP-1 monocytes and CXCL10 mRNA was measured by qRT-PCR TaqMan assay using the ΔΔC t method with 18s as housekeeping gene. For C, data are from three separate experiments and were analyzed by one-way ANOVA with Tukey’s post-hoc test. (D) CXCL10 incubated with GP63 is unable to chemoattract CXCR3+ cells in vitro. Jurkat T cells stably transfected with CXCR3 were seeded on the apical surface of a 5μm transwell insert, with human recombinant CXCL10 pre-incubated with conditioned media from either L. major WT, Δgp63 , or Δgp63 + 1 in the basal chamber. The number of CXCR3+ Jurkats in the basal chamber after 4 hours were counted to assess chemotactic capacity of CXCL10 after exposure to GP63. (E) Proposed model where the host attempts to upregulate CXCL10 in response to infection, but through the activity of GP63 L. major is able to impair signaling through the CXCR3 receptor.

Article Snippet: The CXCL10 plasmid was obtained from Origene (NM_001565), and contains C-terminal Myc and Flag epitope tags.

Techniques: Activity Assay, Infection, Concentration Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, TaqMan Assay, Incubation, In Vitro, Stable Transfection, Transfection, Recombinant

Journal: bioRxiv

Article Title: Pathogen evasion of chemokine response through suppression of CXCL10

doi: 10.1101/557876

Figure Lengend Snippet: (A) LCL 18524 was used to screen L. major (p=0.0001), P. berghei (p=0.99), T. gondii I (RH) (p=0.44), T. gondii II (Prugniaud A7) (p=0.011), S. enterica serovar Typhimurium (p=0.0001), S. aureus (p=0.12), C. trachomatis (p=0.0001), M. marinum (p=0.37), M. smegmatis (p>0.99), and C. neoformans (p=0.010) for CXCL10 suppressing activity (n=2-4 for each pathogen). CXCL10 concentration was measured by ELISA and is represented as the log 2 of fold change relative to uninfected controls. P values calculated by one-way ANOVA with Dunnett’s post-hoc test comparing non-log transformed values to 1, which would represent no change relative to uninfected. (*) represents p<0.01. (B-C) S. Typhimurium and C. trachomatis suppress CXCL10 in a second LCL. Infections were performed in the LCL HG02647 for S. Typhimurium (n=6; two experiments) and C. trachomatis (n=5; three experiments). Mean +/− standard error of the mean is plotted and P values calculated by Student’s t-test . (D) S. Typhimurium suppresses production of CXCL10 in THP-1 monocytes. THP-1 monocytes were stimulated with 1μg/mL of purified LPS from S. Typhimurium at the time of infection. CXCL10 concentration in culture supernatant at 24hpi was assayed by ELISA. Mean +/− standard error the mean is plotted, and P values calculated by two-way ANOVA with Tukey’s post-hoc test. (E) C. trachomatis suppresses CXCL10 in the human endocervical epithelial cell line A2EN. CXCL10 concentration in culture supernatant at 72hpi was assayed by ELISA. Mean +/− standard error of the mean is plotted and P values calculated by Student’s t-test .

Article Snippet: The CXCL10 plasmid was obtained from Origene (NM_001565), and contains C-terminal Myc and Flag epitope tags.

Techniques: Activity Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Transformation Assay, Purification, Infection