Journal: Nature communications

Article Title: A digital single-molecule nanopillar SERS platform for predicting and monitoring immune toxicities in immunotherapy.

doi: 10.1038/s41467-021-21431-w

Figure Lengend Snippet: Fig. 1 Digital single-molecule nanopillar surface-enhanced Raman scattering (SERS) platform for parallel counting of four types of cytokines. SEM images of a pillar array side view, b nanoboxes, and c a single nanobox on the top of a pillar; d SERS spectra of nanoboxes conjugated with 5,5-dithiobis (2-nitrobenzoic acid) (DTNB), 4-mercaptobenzoic acid (MBA), 2,3,5,6-tetrafluoro-4-mercaptobenzoic acid (TFMBA), or 2‐mercapto‐4‐methyl‐5‐ thiazoleacetic acid (MMTAA) Raman reporters; e workflow for multiplex counting of cytokines, including fibroblast growth factor 2 (FGF-2), granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and fractalkine (CX3CL1). Data from one independent experiment.

Article Snippet: FGF-2 (223-FB), G-CSF (214-CS), GM-CSF (215-GM), CX3CL1 (365-FR) cytokines; monoclonal anti-FGF-2 (MAB233), anti-G-CSF (MAB214), anti-GMCSF (MAB615), anti-CX3CL1 (MAB3652) antibodies; polyclonal anti-FGF-2 (AF233), anti-G-CSF (AF-214), anti-GM-CSF (AF-215), anti-CX3CL1 (AF-365) antibodies; and FGF-2 (DY233-05), G-CSF (DY214-05), GM-CSF (DY215-05), and CX3CL1 (DY365) ELISA kits were bought from R&D Systems.

Techniques: Multiplex Assay

Journal: Nature communications

Article Title: A digital single-molecule nanopillar SERS platform for predicting and monitoring immune toxicities in immunotherapy.

doi: 10.1038/s41467-021-21431-w

Figure Lengend Snippet: Fig. 4 Specificity of digital nanopillar SERS platform for FGF-2 cytokine detection. Representative confocal SERS images in the presence of a target FGF-2 (1031 aM), and negative controls with non-target controls b G-CSF (1031 aM), c GM-CSF (1031 aM), d CX3CL1 (1031 aM), and e PBS. The median (interquartile range) of active pillars per scanning image for FGF-2, G-CSF, GM-CSF, CX3CL1, and PBS was 72 (63.5–76.75), 1.5 (1.5–2), 2 (1–4), 0.5 (0–1.25), and 1 (1–1.75), respectively. Data from one independent experiment.

Article Snippet: FGF-2 (223-FB), G-CSF (214-CS), GM-CSF (215-GM), CX3CL1 (365-FR) cytokines; monoclonal anti-FGF-2 (MAB233), anti-G-CSF (MAB214), anti-GMCSF (MAB615), anti-CX3CL1 (MAB3652) antibodies; polyclonal anti-FGF-2 (AF233), anti-G-CSF (AF-214), anti-GM-CSF (AF-215), anti-CX3CL1 (AF-365) antibodies; and FGF-2 (DY233-05), G-CSF (DY214-05), GM-CSF (DY215-05), and CX3CL1 (DY365) ELISA kits were bought from R&D Systems.

Techniques:

Journal: Nature communications

Article Title: A digital single-molecule nanopillar SERS platform for predicting and monitoring immune toxicities in immunotherapy.

doi: 10.1038/s41467-021-21431-w

Figure Lengend Snippet: Fig. 5 Specificity of the digital nanopillar SERS platform for FGF-2 cytokine detection. Representative SEM images of pillar array incubated with FGF-2 SERS nanotags in the presence of a, b FGF-2 (1031 aM), c G-CSF (1031 aM), d GM-CSF (1031 aM), e CX3CL1 (1031 aM), and f PBS. The red circles highlight the existence of SERS nanotags. Panel b is the magnified SEM image of the red-highlighted section in a. It is noted that nanofabrication debris on the sidewall of the pillars can also be seen. Data from one independent experiment.

Article Snippet: FGF-2 (223-FB), G-CSF (214-CS), GM-CSF (215-GM), CX3CL1 (365-FR) cytokines; monoclonal anti-FGF-2 (MAB233), anti-G-CSF (MAB214), anti-GMCSF (MAB615), anti-CX3CL1 (MAB3652) antibodies; polyclonal anti-FGF-2 (AF233), anti-G-CSF (AF-214), anti-GM-CSF (AF-215), anti-CX3CL1 (AF-365) antibodies; and FGF-2 (DY233-05), G-CSF (DY214-05), GM-CSF (DY215-05), and CX3CL1 (DY365) ELISA kits were bought from R&D Systems.

Techniques: Incubation

Journal: Nature communications

Article Title: A digital single-molecule nanopillar SERS platform for predicting and monitoring immune toxicities in immunotherapy.

doi: 10.1038/s41467-021-21431-w

Figure Lengend Snippet: Fig. 6 Sensitivity for the simultaneous detection of four cytokines. Representative confocal SERS images of fibroblast growth factor 2 (FGF-2), granulocyte colony-stimulating factor (GM-CSF), granulocyte colony- stimulating factor (G-CSF), and fractalkine (CX3CL1) with the concentration of a 2.6 aM, b 26 aM, c 260 aM, d 1031 aM. Colour scale bars indicate Raman intensities from 5,5-dithiobis (2-nitrobenzoic acid) (DTNB), 4-mercaptobenzoic acid (MBA), 2,3,5,6-tetrafluoro-4-mercaptobenzoic acid (TFMBA), or 2‐mercapto‐4‐methyl‐5‐thiazoleacetic acid (MMTAA). The median (interquartile range) of active pillars per scanning image of FGF-2, G-CSF, GM-CSF, CX3CL1 for 2.6 aM: 3 (1.5–3), 1 (1–2), 2 (1–3), 2 (1–3); 26 aM: 8 (5.5–10), 10 (9–13), 7 (6–10), 8 (6–10); 260 aM: 40 (36–48), 40 (35–52), 39 (35–50), 37 (36–49); and 1031 aM: 79 (61.5–97), 78 (72–87.5), 88 (68.5–97), 79 (64–95), respectively. Data represents one experiment from three independent tests.

Article Snippet: FGF-2 (223-FB), G-CSF (214-CS), GM-CSF (215-GM), CX3CL1 (365-FR) cytokines; monoclonal anti-FGF-2 (MAB233), anti-G-CSF (MAB214), anti-GMCSF (MAB615), anti-CX3CL1 (MAB3652) antibodies; polyclonal anti-FGF-2 (AF233), anti-G-CSF (AF-214), anti-GM-CSF (AF-215), anti-CX3CL1 (AF-365) antibodies; and FGF-2 (DY233-05), G-CSF (DY214-05), GM-CSF (DY215-05), and CX3CL1 (DY365) ELISA kits were bought from R&D Systems.

Techniques: Concentration Assay

Journal: Nature communications

Article Title: A digital single-molecule nanopillar SERS platform for predicting and monitoring immune toxicities in immunotherapy.

doi: 10.1038/s41467-021-21431-w

Figure Lengend Snippet: Fig. 7 Digital nanopillar SERS assay for monitoring melanoma patients during immune checkpoint therapy. For Patient 1 who developed severe irAEs, SERS images for cytokine detection on a day 7, b day 21, c day 42, d cytokine concentration graph for fibroblast growth factor 2 (FGF-2), granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and fractalkine (CX3CL1). The two shorter horizontal lines denote the interquartile ranges (25th and 75th percentile) and the longer horizontal lines in between denote the median (50th percentile), and e LDA analysis, respectively. For Patient 6 who developed mild irAEs, SERS images for cytokine detection on f day 0, g day 21, h day 42, i four cytokine concentration graph, the two shorter horizontal lines denote the interquartile ranges (25th and 75th percentile) and the longer horizontal lines in between denote the median (50th percentile), and j LDA analysis, respectively. IPI ipilimumab, PEMBRO pembrolizumab; G3 grade 3, G2 grade 2; SD stable disease, PR partial response. For Patient 1, the median (interquartile range) of active pillars per scanning image of FGF-2, G-CSF, GM-CSF, CX3CL1 on day 7: 14 (11–22.5), 23 (21, 29), 12 (7.5–18), 17 (9–25.5); day 21: 30 (19–37.5), 33 (19–41), 26 (17.5–36.5), 29 (21–43); and day 42: 33 (16.5–58.5), 76 (64–128.5), 25 (14–39.5), 48 (26.5–73.5), respectively. For Patient 6, the median (interquartile range) of active pillars per scanning image of FGF-2, G-CSF, GM-CSF, CX3CL1 on day 0: 18 (16–23), 49 (31.5–56), 23 (17.5–28), 20 (14.5–27); day 21: 29 (24–33.5), 53 (46.5–70), 35 (25–46), 22 (19–29.5); and day 42: 13 (8–16.5), 44 (23.5–55.5), 10 (6.5–12.5), 30 (24–34.5), respectively. The data represented three technical replicates obtained from three chips. Nine images were acquired from each chip for cytokine counting. Statistical analysis was based on Kruskal–Wallis test followed by Dunn’s test to correct multiple comparisons (two-sided). Source data are provided in the Source Data file.

Article Snippet: FGF-2 (223-FB), G-CSF (214-CS), GM-CSF (215-GM), CX3CL1 (365-FR) cytokines; monoclonal anti-FGF-2 (MAB233), anti-G-CSF (MAB214), anti-GMCSF (MAB615), anti-CX3CL1 (MAB3652) antibodies; polyclonal anti-FGF-2 (AF233), anti-G-CSF (AF-214), anti-GM-CSF (AF-215), anti-CX3CL1 (AF-365) antibodies; and FGF-2 (DY233-05), G-CSF (DY214-05), GM-CSF (DY215-05), and CX3CL1 (DY365) ELISA kits were bought from R&D Systems.

Techniques: Concentration Assay

Journal: Mediators of inflammation

Article Title: Growth Arrest-Specific 6 Enhances the Suppressive Function of CD4 + CD25 + Regulatory T Cells Mainly through Axl Receptor.

doi: 10.1155/2017/6848430

Figure Lengend Snippet: Figure 6: Gas6 modulates the Foxp3 and CTLA4 expression mainly through Axl receptor. (a–d) CD4+CD25+Tregs were treated with anti- Axl or anti-Mertk Abs or PBS in the presence of 100 ng/ml rmGas6. After 24 h of incubation, the expression of CTLA-4 and Foxp3 was determined by flow cytometry (𝑛= 4/group). ∗𝑃< 0.05 compared with the value for the Gas6 group, and #𝑃< 0.05 compared with the value for rmGas6+anti-Mertk group. (e–h) The expression of CTLA-4 and Foxp3 in Tregs with or without Axl knockout was determined by flow cytometry. ∗𝑃< 0.05 compared with the value for the Gas6 group.

Article Snippet: To investigate the effect of Gas6 on CD4+CD25+Tregs in vivo, healthy mice were administered 1, 3, or 6 μg/mouse of rmGas6 (8310-GS, R&D Systems, Minneapolis, MN) via tail vein.

Techniques: Expressing, Incubation, Flow Cytometry, Knock-Out

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 1. Mean (±SEM) microsphere- derived risk area from animals assigned to either (A) 30 min or (B) 45 min of coronary occlusion. C, Mean (±SEM) microvascular blood flow from myocardial contrast echocardiography (MCE) performed during coronary occlusion in the risk area and remote territories, as well as from sham-treated mice. D, Example of background-subtracted, color-coded MCE images during coronary occlusion, and time-intensity data after a destructive pulse sequence from the risk area and remote territory. For images, time after destructive pulse is at upper left and color-coded scale at bottom of the 5 s image. ADAMTS13 indicates a disintegrin and metalloprotein- ase with a thrombospondin type-1 motif member 13; LV, left ventricle; RA, risk area; and WT, wild-type. *P <0.05 vs remote territory.

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Derivative Assay, Sequencing

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 2. Postischemic myocardial perfusion by MCE in mice undergo- ing 30 min of ischemia and reperfusion measured in the remote region and the risk area. Data (mean±SEM) are shown for (A) myocardial microvascular blood flow (MBF), (B) microvascular blood volume (MBV), and (C) microvascular flux rate (β). ADAMTS13 indicates a disintegrin and metalloproteinase with a thrombo- spondin type-1 motif member 13. *P <0.05 vs remote territory; †P <0.05 vs WT (wild type); ‡P <0.05 only before correction for multiple comparison.

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Comparison

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 3. Mean (±SEM) signal enhance- ment on MCE molecular imaging after 30 min of ischemia and reperfusion for (A) VWF (von Willebrand factor) A-1 domain and (B) platelet GP (glycoprotein) Ibɑ. Data are expressed as signal difference between targeted and control microbubbles (MB). C, Examples of color-coded MCE molecular imaging in the short-axis plane for targeted and control MBs (color scale at bottom) and the microsphere-derived risk area in the same short-axis plane. *P <0.05 vs remote territory; †P <0.05 vs wild-type and ADAMTS13−/− (a disintegrin and metal- loproteinase with a thrombospondin type-1 motif member 13); ‡P <0.05 vs WT (wild type).

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Imaging, Control, Derivative Assay

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 5. Mean (±SEM) infarct size by triphenyltetrazolium chloride staining as a percentage of the total microsphere-de- rived risk area for mice undergoing either (A) 30 min of ischemia followed by 90 min of reperfusion or (B) 45 min of ischemia followed by 3 d of reperfusion. *P <0.05 vs WT (wild type) before Bonfer- roni correction; †P <0.05 vs WT+ADAMTS13 (a disintegrin and metalloproteinase with a thrombospondin type-1 motif member 13) after Bonferroni correction.

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Staining

Journal: Circulation: Cardiovascular Imaging

Article Title: Molecular Imaging of VWF (von Willebrand Factor) and Platelet Adhesion in Postischemic Impaired Microvascular Reflow

doi: 10.1161/circimaging.118.007913

Figure Lengend Snippet: Figure 4. Examples of immunohistochemis- try from the sham-treated control WT (wild type) mice (anterior and posterior myocar- dial with anterior labeled as risk area) and from postischemic WT and ADAMTS13−/− (a disintegrin and metalloproteinase with a thrombospondin type-1 motif member 13) mice. The top and bottom rows show fused im- ages for platelet CD41 immunostaining (red), endothelial lectin staining (green), and nuclear counterstain (blue). The middle row shows corresponding images for only the red channel (platelet CD41) in the risk area. For the insets, arrowheads show single platelets, whereas the arrow in the ADAMTS13−/− mouse illustrates 3 platelets linearly arranged (scale bar=20 μm). MI indicates myocardial infarction.

Article Snippet: In Vivo Imaging Study Design Protocol 1 Closed-chest myocardial IR with 30 minutes of transient left anterior descending coronary artery (LAD) occlusion was performed in WT mice (n=40), WT mice treated with recombinant full-length human ADAMTS13, 5 μg IV (R&D Systems, Minneapolis, MN) just before IR (n=14), and ADAMTS13−/− mice (n=16).

Techniques: Control, Labeling, Immunostaining, Staining

Journal: Journal of pharmacological sciences

Article Title: Lymphangiogenesis induced by vascular endothelial growth factor receptor 1 signaling contributes to the progression of endometriosis in mice.

doi: 10.1016/j.jphs.2020.05.003

Figure Lengend Snippet: Fig. 3. Accumulation of macrophages and fibroblasts in endometrial implant tissues. (A) The levels of the mRNAs encoding the lymphangiogenic growth factors VEGF-C and VEGF-D in endometrial tissues from WT/WT and TK-/-/TK-/- mice. Data are expressed as the mean ± SEM (n ¼ 5e6 mice per group). *P < 0.05. (B) Double immunostaining of VEGF-C (green) or VEGF-D (green), and CD11b (red) or S100A4 (red) in endometrial tissues from WT/WT mice at Day 14. Scale bars, 50 mm. (C) Left: Immunostaining of CD11b (green) in endometrial tissues from WT/WT and TK-/-/TK-/- mice at Day 14. Scale bars, 100 mm. Right: The numbers of CD11bþ cells in endometrial tissues from WT/WT and TK-/-/TK-/-

Article Snippet: The sections were incubated with the following primary antibodies at 4 C overnight: a rabbit anti-mouse VEGFR1 polyclonal antibody (1:200; Abcam), a goat anti-mouse VEGFR1 polyclonal antibody (1:200; Santa Cruz Biotechnology, Santa Cruz, CA, USA), a rabbit anti-LYVE-1 polyclonal antibody (1:200; Abcam), a goat anti-LYVE-1 polyclonal antibody (1:200; R&D Systems Inc., Minneapolis, MN, USA), a rabbit anti-VEGF-C polyclonal antibody (1:200; Abcam), a goat anti-VEGFD polyclonal antibody (1:40; R&D Systems Inc.), a rat anti-mouse CD11b monoclonal antibody (1:200; BD Biosciences, San Jose, CA, USA), a rabbit anti-mouse S100A4 polyclonal antibody (1:200; Abcam), or a goat anti-mouse S100A4 polyclonal antibody (1:100; OriGene Technologies, Rockville, MD, USA).

Techniques: Double Immunostaining, Immunostaining