Journal: International Journal of Molecular Sciences

Article Title: Possible Roles of CC- and CXC-Chemokines in Regulating Bovine Endometrial Function during Early Pregnancy

doi: 10.3390/ijms18040742

Figure Lengend Snippet: Comparison of mRNA levels for selected chemokines in the endometrium of pregnant vs. non-pregnant cows as determined by microarray analysis (Fold > 2.0; p < 0.05).

Article Snippet: Cultured endometrial tissues were further incubated in the medium with recombinant proteins as follows: bovine CCL2 (RP0027B, Kingfisher Biotech., Inc. St. Paul, MN, USA), human CCL8 (281-CP, R&D Systems, Inc. Minneapolis, MN, USA), bovine CCL11 (RP0071B, Kingfisher Biotech.), human CCL14 (1578-HC, R&D Systems), human CCL16 (TP723266, OriGene Technologies, Inc., Rockville, MD, USA), bovine CXCL10 (RP0079B, Kingfisher Biotech.), bovine tumor necrosis factor-α (TNF; 2279-BT, R&D Systems), human leukemia inhibitory factor (LIF; TP723270, OriGene Technologies), bovine IFNT (1.1 × 10 5 U/mg, generated from HEK293 cells as described previously; Takahashi et al., 2017 [ ]) or supernatant derived from homogenized fetal trophoblast on day 18 of pregnancy (FMP).

Techniques: Comparison, Microarray

Journal: International Journal of Molecular Sciences

Article Title: Possible Roles of CC- and CXC-Chemokines in Regulating Bovine Endometrial Function during Early Pregnancy

doi: 10.3390/ijms18040742

Figure Lengend Snippet: Changes in relative amounts of mRNA for ( a ) CCL2, ( b ) CCL8, ( c ) CCL11, ( d ) CCL14, ( e ) CCL16, and ( f ) CXCL10 in the endometrium at days 15 and 18 of non-pregnant cows (NP) and pregnant cows (P). Data are means ± SEM of four cows per stage and are expressed as relative ratios of the mRNAs to SUZ12 polycomb repressive complex 2 subunit (SUZ12). p -Values show significant differences between NP and P.

Article Snippet: Cultured endometrial tissues were further incubated in the medium with recombinant proteins as follows: bovine CCL2 (RP0027B, Kingfisher Biotech., Inc. St. Paul, MN, USA), human CCL8 (281-CP, R&D Systems, Inc. Minneapolis, MN, USA), bovine CCL11 (RP0071B, Kingfisher Biotech.), human CCL14 (1578-HC, R&D Systems), human CCL16 (TP723266, OriGene Technologies, Inc., Rockville, MD, USA), bovine CXCL10 (RP0079B, Kingfisher Biotech.), bovine tumor necrosis factor-α (TNF; 2279-BT, R&D Systems), human leukemia inhibitory factor (LIF; TP723270, OriGene Technologies), bovine IFNT (1.1 × 10 5 U/mg, generated from HEK293 cells as described previously; Takahashi et al., 2017 [ ]) or supernatant derived from homogenized fetal trophoblast on day 18 of pregnancy (FMP).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Possible Roles of CC- and CXC-Chemokines in Regulating Bovine Endometrial Function during Early Pregnancy

doi: 10.3390/ijms18040742

Figure Lengend Snippet: Localization of CCR1 (binds to CCL8, CCL14, and CCL16), CCR2 (binds to CCL2, CCL8, and CCL16), CCR3 (binds to CCL11), and CXCR3 (binds to CXCL10) in the bovine endometrium and fetal trophoblast obtained from cows in their 18th day of pregnancy. Intensive immunoreactivity was observed in endometrial epithelial cells, glandular epithelial cells, or fetal trophoblast. No positive immunoreactivity was observed in the negative control (Control). Scale bar = 50 µm.

Article Snippet: Cultured endometrial tissues were further incubated in the medium with recombinant proteins as follows: bovine CCL2 (RP0027B, Kingfisher Biotech., Inc. St. Paul, MN, USA), human CCL8 (281-CP, R&D Systems, Inc. Minneapolis, MN, USA), bovine CCL11 (RP0071B, Kingfisher Biotech.), human CCL14 (1578-HC, R&D Systems), human CCL16 (TP723266, OriGene Technologies, Inc., Rockville, MD, USA), bovine CXCL10 (RP0079B, Kingfisher Biotech.), bovine tumor necrosis factor-α (TNF; 2279-BT, R&D Systems), human leukemia inhibitory factor (LIF; TP723270, OriGene Technologies), bovine IFNT (1.1 × 10 5 U/mg, generated from HEK293 cells as described previously; Takahashi et al., 2017 [ ]) or supernatant derived from homogenized fetal trophoblast on day 18 of pregnancy (FMP).

Techniques: Negative Control, Control

Journal: International Journal of Molecular Sciences

Article Title: Possible Roles of CC- and CXC-Chemokines in Regulating Bovine Endometrial Function during Early Pregnancy

doi: 10.3390/ijms18040742

Figure Lengend Snippet: Effects of the supernatant derived from homogenized fetal trophoblast (FMP; 200 ng/mL) and interferon-τ (IFNT; 100 ng/mL) on the mRNA expression of ( a ) CCL2, ( b ) CCL8, ( c ) CCL11, ( d ) CCL14, ( e ) CCL16, and ( f ) CXCL10 in cultured bovine endometrial tissues. Homogenization buffer was added at the control group. Data are means ± SEM of five cows and are expressed as relative ratios of the mRNAs to SUZ12. p -Values show significant differences between treated group and control group.

Article Snippet: Cultured endometrial tissues were further incubated in the medium with recombinant proteins as follows: bovine CCL2 (RP0027B, Kingfisher Biotech., Inc. St. Paul, MN, USA), human CCL8 (281-CP, R&D Systems, Inc. Minneapolis, MN, USA), bovine CCL11 (RP0071B, Kingfisher Biotech.), human CCL14 (1578-HC, R&D Systems), human CCL16 (TP723266, OriGene Technologies, Inc., Rockville, MD, USA), bovine CXCL10 (RP0079B, Kingfisher Biotech.), bovine tumor necrosis factor-α (TNF; 2279-BT, R&D Systems), human leukemia inhibitory factor (LIF; TP723270, OriGene Technologies), bovine IFNT (1.1 × 10 5 U/mg, generated from HEK293 cells as described previously; Takahashi et al., 2017 [ ]) or supernatant derived from homogenized fetal trophoblast on day 18 of pregnancy (FMP).

Techniques: Derivative Assay, Expressing, Cell Culture, Homogenization, Control

Journal: International Journal of Molecular Sciences

Article Title: Possible Roles of CC- and CXC-Chemokines in Regulating Bovine Endometrial Function during Early Pregnancy

doi: 10.3390/ijms18040742

Figure Lengend Snippet: Effects of CCL2, CCL8, CCL11, CCL14, CCL16, and CXCL10 (50 ng/mL each) on the mRNA expression of ( a ) interferon-stimulated gene 15 (ISG15), ( b ) myxovirus-resistance gene 1 (MX1), ( c ) cyclooxygenase 2 (COX2), ( d ) oxytocin receptor (OTR), and ( e ) estrogen receptor α (ESR1) in cultured bovine endometrial tissues. Data are means ± SEM of five cows and are expressed as relative ratios of the mRNAs to SUZ12. p -Values show significant differences between treated group and control group.

Article Snippet: Cultured endometrial tissues were further incubated in the medium with recombinant proteins as follows: bovine CCL2 (RP0027B, Kingfisher Biotech., Inc. St. Paul, MN, USA), human CCL8 (281-CP, R&D Systems, Inc. Minneapolis, MN, USA), bovine CCL11 (RP0071B, Kingfisher Biotech.), human CCL14 (1578-HC, R&D Systems), human CCL16 (TP723266, OriGene Technologies, Inc., Rockville, MD, USA), bovine CXCL10 (RP0079B, Kingfisher Biotech.), bovine tumor necrosis factor-α (TNF; 2279-BT, R&D Systems), human leukemia inhibitory factor (LIF; TP723270, OriGene Technologies), bovine IFNT (1.1 × 10 5 U/mg, generated from HEK293 cells as described previously; Takahashi et al., 2017 [ ]) or supernatant derived from homogenized fetal trophoblast on day 18 of pregnancy (FMP).

Techniques: Expressing, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Possible Roles of CC- and CXC-Chemokines in Regulating Bovine Endometrial Function during Early Pregnancy

doi: 10.3390/ijms18040742

Figure Lengend Snippet: Primers used in real-time PCR.

Article Snippet: Cultured endometrial tissues were further incubated in the medium with recombinant proteins as follows: bovine CCL2 (RP0027B, Kingfisher Biotech., Inc. St. Paul, MN, USA), human CCL8 (281-CP, R&D Systems, Inc. Minneapolis, MN, USA), bovine CCL11 (RP0071B, Kingfisher Biotech.), human CCL14 (1578-HC, R&D Systems), human CCL16 (TP723266, OriGene Technologies, Inc., Rockville, MD, USA), bovine CXCL10 (RP0079B, Kingfisher Biotech.), bovine tumor necrosis factor-α (TNF; 2279-BT, R&D Systems), human leukemia inhibitory factor (LIF; TP723270, OriGene Technologies), bovine IFNT (1.1 × 10 5 U/mg, generated from HEK293 cells as described previously; Takahashi et al., 2017 [ ]) or supernatant derived from homogenized fetal trophoblast on day 18 of pregnancy (FMP).

Techniques: Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Possible Roles of CC- and CXC-Chemokines in Regulating Bovine Endometrial Function during Early Pregnancy

doi: 10.3390/ijms18040742

Figure Lengend Snippet: Hypothetical model for inhibition of luteolysis by IFNT and chemokines. Although this model is not concerned with the effects of steroids or growth factors, IFNT, CCL2, CCL8, CCL16, CXCL10, and LIF may block TNF-stimulated-COX2 expression in bovine endometrial cells, leading to the reduction of TNF-induced PGF2α output from the cells. Furthermore, IFNT and CCL16 may stimulate anti-viral activity by up-regulating ISG15 and MX1 expression at the time of maternal recognition in cows. Red and blue arrows show stimulatory and inhibitory actions of each substance, respectively. IFNT may stimulate both CCL8 and CXCL10 production and inhibit CCL14 production from bovine endometrium. Effects of CCL11 on bovine endometrial function are still unclear, although its receptor (CCR3) is expressed in the endometrial epithelial cells.

Article Snippet: Cultured endometrial tissues were further incubated in the medium with recombinant proteins as follows: bovine CCL2 (RP0027B, Kingfisher Biotech., Inc. St. Paul, MN, USA), human CCL8 (281-CP, R&D Systems, Inc. Minneapolis, MN, USA), bovine CCL11 (RP0071B, Kingfisher Biotech.), human CCL14 (1578-HC, R&D Systems), human CCL16 (TP723266, OriGene Technologies, Inc., Rockville, MD, USA), bovine CXCL10 (RP0079B, Kingfisher Biotech.), bovine tumor necrosis factor-α (TNF; 2279-BT, R&D Systems), human leukemia inhibitory factor (LIF; TP723270, OriGene Technologies), bovine IFNT (1.1 × 10 5 U/mg, generated from HEK293 cells as described previously; Takahashi et al., 2017 [ ]) or supernatant derived from homogenized fetal trophoblast on day 18 of pregnancy (FMP).

Techniques: Inhibition, Blocking Assay, Expressing, Activity Assay

Journal: International journal of cancer

Article Title: Tumor-stromal crosstalk in invasion of oral squamous cell carcinoma: a pivotal role of CCL7.

doi: 10.1002/ijc.25060

Figure Lengend Snippet: Figure 1. CAF induces invasive growth of cancer cells, and CCL7 mRNA expression is increased in CAF by coculturing with OSCC cells. (a)

Article Snippet: Human recombinant monocyte chemotactic protein CCL7, human recombinant IL-1a and VEGF were obtained from R&D Systems (R&D, Minneapolis, MN).

Techniques: Expressing

Journal: International journal of cancer

Article Title: Tumor-stromal crosstalk in invasion of oral squamous cell carcinoma: a pivotal role of CCL7.

doi: 10.1002/ijc.25060

Figure Lengend Snippet: Figure 2. CAF dramatically increased the release of CCL7 by coculturing with OSCC cells. (a) Five OSCC cell lines (YD-10B, HSC-2, HSC-3,

Article Snippet: Human recombinant monocyte chemotactic protein CCL7, human recombinant IL-1a and VEGF were obtained from R&D Systems (R&D, Minneapolis, MN).

Techniques:

Journal: International journal of cancer

Article Title: Tumor-stromal crosstalk in invasion of oral squamous cell carcinoma: a pivotal role of CCL7.

doi: 10.1002/ijc.25060

Figure Lengend Snippet: Figure 3. CCL7 promoted OSCC cell migration and induced membrane ruffling and cell spreading in OSCC cells. (a) Confluent monolayers of

Article Snippet: Human recombinant monocyte chemotactic protein CCL7, human recombinant IL-1a and VEGF were obtained from R&D Systems (R&D, Minneapolis, MN).

Techniques: Migration, Membrane

Journal: International journal of cancer

Article Title: Tumor-stromal crosstalk in invasion of oral squamous cell carcinoma: a pivotal role of CCL7.

doi: 10.1002/ijc.25060

Figure Lengend Snippet: Figure 4. Invasion of OSCC cells was inhibited by pretreatment with CCL7 neutralizing antibody. (a) YD-10B or HSC-2 cells in serum-free media were placed in the upper well of a 24-transwell plate with collagen-coated filters (8 lm pore). CAF-CM was added into the lower well to induce

Article Snippet: Human recombinant monocyte chemotactic protein CCL7, human recombinant IL-1a and VEGF were obtained from R&D Systems (R&D, Minneapolis, MN).

Techniques:

Journal: International journal of cancer

Article Title: Tumor-stromal crosstalk in invasion of oral squamous cell carcinoma: a pivotal role of CCL7.

doi: 10.1002/ijc.25060

Figure Lengend Snippet: Figure 5. Receptors for CCL7 in OSCC cells were shown to be CCR1 and CCR3. (a) Expression of CCR1, CCR2 and CCR3 in OSCC cells, and

Article Snippet: Human recombinant monocyte chemotactic protein CCL7, human recombinant IL-1a and VEGF were obtained from R&D Systems (R&D, Minneapolis, MN).

Techniques: Expressing

Journal: International journal of cancer

Article Title: Tumor-stromal crosstalk in invasion of oral squamous cell carcinoma: a pivotal role of CCL7.

doi: 10.1002/ijc.25060

Figure Lengend Snippet: Figure 6. IL-1a increased CCL7 secretion by CAF. (a) Culture medium collected from Ca9.22, which induces the highest level of CCL7

Article Snippet: Human recombinant monocyte chemotactic protein CCL7, human recombinant IL-1a and VEGF were obtained from R&D Systems (R&D, Minneapolis, MN).

Techniques:

Journal: eBioMedicine

Article Title: Soluble CD46 as a diagnostic marker of hepatic steatosis

doi: 10.1016/j.ebiom.2024.105184

Figure Lengend Snippet: Discovery of soluble CD46 as a marker of steatotic human hepatocytes. ( a ) Frequency of intrahepatic IL-4 + iNKT cells with respect to iNKT cells isolated from normal human liver tissue [Grade 0, n = 26; Grade ≥1, n = 11; Wilcoxon rank-sum test after Shapiro–Wilk test was significant (p = 0.0336)]. ( b ) Frequency of IL-4 + iNKT cells after 7 days’ expansion, alone or in coculture with unloaded (UL)- or fat-loaded (FL)- HepaRG cells [n = 8; one-way repeated measures ANOVA p < 0.0001; post-hoc paired t-test with BH-adjusted p-values]. ( c ) Flow cytometric expression analysis of 369 surface markers on paired UL- and FL-HepaRG cell cultures [n = 6; paired t-test with BH-adjusted p-values]. ( d ) Concentration of sCD46 in supernatants from UL- and FL-HepaRG cultures measured with a competitive cell-based flow cytometry assay [n = 20; unpaired t-test]. ( e ) Concentration of sCD46 in supernatants from UL- and FL-primary human liver organoid cultures measured by ELISA [n = 8; paired t-test]. ( f ) Frequency of IL-4 + iNKT cells after 7 days’ expansion in the absence or presence of recombinant human CD46-Fc or control Fc protein [n = 6, one-way repeated measures ANOVA p = 0.0008; post-hoc paired t-test with BH-adjusted p-values]. ( g ) Frequency of IL-4 + iNKT cells after 7 days’ expansion in UL-HepaRG cell cocultures treated with of 5 μg/ml neutralizing α-CD46 or isotype-control antibody [n = 14; paired t-test]. ( h ) Frequency of IL-4 + iNKT cells after 7 days’ expansion in coculture with UL-HepaRG cells stably transfected with random-sequence shRNA or CD46 shRNA. Untransfected UL-HepaRG cells were used as controls [n = 15; one-way repeated measures ANOVA p = 0.0007; post-hoc paired t-test with BH-adjusted p-values]. ( i ) Frequency of IL-4 + iNKT cells after 7 days’ expansion in coculture with UL- or FL-HepaRG cells that were untreated or treated with 10 μM MMP inhibitor (TAPI-1, GI254023X and Batimastat) or DMSO [n = 8; two-way repeated measures ANOVA (p treatment < 0.0001, p loading = 0.043); post-hoc paired t-test with BH-adjusted p-values].

Article Snippet: Treatment of ex vivo -expanded iNKT cells with chimeric CD46-Fc (10257-CD; R&D Systems) or control Fc protein was renewed with each medium change.

Techniques: Marker, Isolation, Expressing, Concentration Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Recombinant, Control, Stable Transfection, Transfection, Sequencing, shRNA

Journal: bioRxiv

Article Title: Chemokine profiling of melanoma-macrophage crosstalk identifies CCL8 and CCL15 as prognostic factors in cutaneous melanoma

doi: 10.1101/2023.10.04.560856

Figure Lengend Snippet: M1 and M2 macrophages were co-cultured with BLM melanoma cells for transcriptomic (24 h) and secretomic (72 h) analyses. (A) Gene set enrichment analyses (GSEA) comparing 24 hours exposed vs un-exposed isolated cells, as indicated (n=3 donors). Normalized Enrichment Score (NES) values from the main upregulated hallmark_pathwaysfor all groups,are shown. GOMF and KEGG gene sets were additionally studied for cytokine/ chemokine inflammatory pathways. False Discovery Rates are represented (FDR q value <0.01). (B) Representative scheme highlighting the coincident most upregulated secreted chemokines (fold >4 to both unex-posed macrophages and melanoma cells). (C) Secretomic analysis (38 chemokines) of 72 hours co-cultured supernatants comparing M1+ BLM or M2+BLM with unexposed macrophages (x-axis) or BLM (y-axis) cells. Fold logarithmic average changes are shown (n=4 donors). (D) CCL8 and CCL15 secretion assessed at 72 h macrophages ±BLM conditioned media. Mean ±standard deviation (SD) values are shown (n= 5 do-nors). (E) CCL8 and CCL15 mRNA expression levels in mutually conditioned isolated cells. Mean ±SD values relative to TBP mRNA are shown (n= 4). Significant differences to the respective untreated controls, are shown (*p< 0.05, unpaired t-test).

Article Snippet: Serum-deprived media ±recombinant human CCL8 or CCL15 (R&D Systems, Minnneapolis, MN) and ±5 μg/ml neutralizing antibodies for CCR1, CCR3 and CCR5 (supplementary material, Table S1) were added on top of the gels, and spheroids were allowed to invade for 72 hours.

Techniques: Cell Culture, Isolation, Standard Deviation, Expressing

Journal: bioRxiv

Article Title: Chemokine profiling of melanoma-macrophage crosstalk identifies CCL8 and CCL15 as prognostic factors in cutaneous melanoma

doi: 10.1101/2023.10.04.560856

Figure Lengend Snippet: (A) Serum deprivation survival assays of BLM, A375 and Skmel-103 melanoma cell lines in response to increasing concentra-tions of CCL8 and CCL15 chemokines (ng/ml), and to FCS 1%, as positive control. Mean ±SD optical density (OD) values, are shown (n= 4). (B) Proliferation response of melanoma cell lines to 200 ng/ml exogenous CCL8 and CCL15, and to FCS 1% as positive control. Mean ±SD percentages of BrdU+ nuclei, are shown (n= 3). (C) BLM spheroids embedded in 3D-collagen and allowed to invade for 72 h in the presence of increasing concentrations of exogenous CCL8 and CCL15, as indicated (ng/ml). Functional chemotactic axes were identified by using neutralizing antibodies against CCR1, CCR3, CCR5 and their corresponding control isotypes (5 μg/ml). Mean ±SD fold-invasion values, are shown (n= 3-6). Images show representative collagen-embedded spheroids showing basal and CCL15 stimulated invasion. Scale bar, 150 μm. (D) Immunoblots showing CCR1, CCR3 and CCR5 expression in whole-lysates of three melanoma cell lines and isolated CD14+ monocytes. (E) Serum deprivation survival and 3D-collagen invasion assays of monocytes in response to 100 ng/ml exogenous chemokines, including FCS 1%, as positive control (n= 5 donors). Significant differences to the respective untreated controls, are shown (*p< 0.05, paired t-test).

Article Snippet: Serum-deprived media ±recombinant human CCL8 or CCL15 (R&D Systems, Minnneapolis, MN) and ±5 μg/ml neutralizing antibodies for CCR1, CCR3 and CCR5 (supplementary material, Table S1) were added on top of the gels, and spheroids were allowed to invade for 72 hours.

Techniques: Positive Control, Functional Assay, Control, Western Blot, Expressing, Isolation

Journal: bioRxiv

Article Title: Chemokine profiling of melanoma-macrophage crosstalk identifies CCL8 and CCL15 as prognostic factors in cutaneous melanoma

doi: 10.1101/2023.10.04.560856

Figure Lengend Snippet: TAM (A) and tumor cell (B) average MFI of CCL15, CCL8, CCR1, CCR3 and CCR5 in non-metastasizing and metastasizing primary tumors.Mann-Whitney statistical analysis was used to compare non-metastasizing vs metastasizing melanomas; p-values are shown.(C) FFPE human melanoma samples stained for CD68 (TAM marker, red) and CCL15, CCL8, CCR1, CCR3 or CCR5 (green). Scale bar, 50 μm.

Article Snippet: Serum-deprived media ±recombinant human CCL8 or CCL15 (R&D Systems, Minnneapolis, MN) and ±5 μg/ml neutralizing antibodies for CCR1, CCR3 and CCR5 (supplementary material, Table S1) were added on top of the gels, and spheroids were allowed to invade for 72 hours.

Techniques: MANN-WHITNEY, Staining, Marker

Journal: bioRxiv

Article Title: Chemokine profiling of melanoma-macrophage crosstalk identifies CCL8 and CCL15 as prognostic factors in cutaneous melanoma

doi: 10.1101/2023.10.04.560856

Figure Lengend Snippet: (A)Disease-free and overall survival 10-year Kaplan–Meier curves. Youden’s index was used to choose a cutoff point to classify the 67 primary melanomas as ‘low’ or ‘high’: TC CCL8 (MFI, 80 a.u.), TC CCL15 (98a.u.),TAM CCL15 (69 a.u.) and TAM CCR3 (78 a.u.). Log-rank p values, are shown. (B) Summarizing model, created with BioRender.com .

Article Snippet: Serum-deprived media ±recombinant human CCL8 or CCL15 (R&D Systems, Minnneapolis, MN) and ±5 μg/ml neutralizing antibodies for CCR1, CCR3 and CCR5 (supplementary material, Table S1) were added on top of the gels, and spheroids were allowed to invade for 72 hours.

Techniques: