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miRNAs regulate the balance of classical and alternative macrophage activat...
Published: 15 February 2023
FIGURE 1. miRNAs regulate the balance of classical and alternative macrophage activation. ( A ) A group of miRNAs enhance “M1-like” macrophage activation by promoting cell signaling, cytokine production, and proliferation through targeting key inhibitors these pathways with these cells (left). A group of miRNAs can also help to dampen or resolve inflammation by restricting or decreasing macrophage proinflammatory activation (right). ( B ) miRNAs can augment alternative macrophage activation by inhibiting M1-like pathways and targeting genes that normally limit M2-like programs (left). Other miRNAs can inhibit alternative macrophage activation by targeting key signaling molecules in this pathway (right). LTB4, leukotriene B4; MAPK, mitogen-activated protein kinase; PGE2, prostaglandin E2. More
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miRNAs regulate metabolic and LPS tolerization programs. ( A ) miRNAs regul...
Published: 15 February 2023
FIGURE 2. miRNAs regulate metabolic and LPS tolerization programs. ( A ) miRNAs regulate multiple facets of lipid metabolism within macrophages. ( B ) miRNAs regulate tolerization through controlling sensitivity to signaling and epigenetic reprograming of the cell. RISC, RNA-induced silencing comp... More
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Published: 15 February 2023
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Published: 15 February 2023
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Published: 15 February 2023
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Published: 15 February 2023
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Deletion of <em>Jag1</em> in DCs.   ( A ) Histograms of JAG1 staini...
Published: 02 February 2023
FIGURE 1. Deletion of Jag1 in DCs. ( A ) Histograms of JAG1 staining in lung CD11c+CD11b+ cells using samples from unsensitized mice and mice that had been sensitized and challenged with OVA. ( B ) Quantification of (A) for the indicated cell subsets in the lungs of unsensitized mice. *p = 0.04, n = 3 mice per group. ( C ) Proportion of JAG1+ cells in the spleens of Jagged-1ffCD11cCre+ animals or controls prior to sensitization with OVA. *p = 0.04, n = 3 mice per group. ( D ) Expression of exon 2 of Jag1 expressed as a fold increase over Jagged-1ffCD11cCre+ in CD11c+CD11b+ cells sorted from the lung 48 h after OVA challenge. *p = 0.1, n = 3 mice per group. Significance in this figure was determined by an unpaired t test. More
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<em>Jag1</em> deletion increases Th2 responses.   ( A ) Levels of c...
Published: 02 February 2023
FIGURE 2. Jag1 deletion increases Th2 responses. ( A ) Levels of cytokines in cell culture supernatants of lung-draining lymph nodes stimulated with OVA for 48 h. *p ≤ 0.01 as determined by an unpaired t test. ( B ) Expression of Muc5ac in lungs of mice sensitized and challenged with OVA as determined by qPCR. *p = 0.03 as determined by two-way ANOVA, with genotype accounting for 14% of the variance in the experiment. ( C ) Number of eosinophils in mice from (B) using whole lung at 48 h after OVA challenge as determined by flow cytometry for Siglec-F+CD11b+MHC class II cells. *p = 0.04 using an unpaired t test. ( D ) Micrographs of lung tissue formalin-fixed, paraffin-embedded sections stained with periodic acid–Schiff (PAS; original magnification, ×200) and H&E (original magnification, ×400) highlighting mucus production and leukocyte infiltration in this model. More
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JAG1 alters Notch 1 receptor expression.   ( A ) qPCR for the <em>Notch</em>...
Published: 02 February 2023
FIGURE 3. JAG1 alters Notch 1 receptor expression. ( A ) qPCR for the Notch1 receptor gene demonstrates a significant reduction in gene expression at 4 h after stimulation with anti-CD3/anti-CD28 in the presence of 5 nM rJAG1. *p < 0.0001 as determined by two-way ANOVA with Bonferroni’s posttest; n = 4 samples per group. ( B ) Significant reduction in surface receptor Notch 1, as determined by flow cytometry, after stimulation of CD4+ T cells with anti-CD3/anti-CD28 for 24 h in the presence of 10 nM rJAG1, with no change in T cell viability ( C ) under the same conditions. *p < 0.0001 using a one-way ANOVA with a posttest. ( D and E ) MFI of Notch 1 in CD4+ T cell subsets in the lung and lung-draining lymph nodes at 48 h after OVA challenge. *p < 0.01, **p < 0.001, ***p < 0.0001 using an unpaired t test. ( F ) Histogram of data quantified in (E), focused on the memory cell subset. ( G ) Quantification of the percentage of each CD4+ T cell subset that was expressing Notch receptor at 48 h after OVA challenge. ***p < 0.0001 as determined by an unpaired t test. For (D), (E), and (G), n = 6 mice per group. More
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Expression of Notch receptors on OT-II<sup>+</sup> cells.   ( A ) Schematic...
Published: 02 February 2023
FIGURE 4. Expression of Notch receptors on OT-II+ cells. ( A ) Schematic of the transfer model used. ( B ) Intracellular staining for IL-4 after stimulation of splenocytes from mice sensitized with OVA after in vitro stimulation with either whole OVA or the OVA peptide specific for the OT-II receptor in the presence of anti-JAG1 or isotype control. ( C ) MFI of Notch 1 and Notch 2 on CD45.1+CD4+CD44hi host cells and CD45.2+CD4+ OT-II cells in the lung and lung-draining lymph node. ( D ) Percentage of CD45.1+ CD4+CD44hi host cells and CD45.2+ OT-II cells expressing Notch 1 and Notch 2 in the spleen. ( E ) Flow cytometry plots of Notch 1 receptor expression on CD4+CD44hi splenocytes after sensitization with OVA for 6 wk. More
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Effect of JAG1 blockade on expansion of CD4<sup>+</sup> T cell subsets.   (...
Published: 02 February 2023
FIGURE 5. Effect of JAG1 blockade on expansion of CD4+ T cell subsets. ( A ) Schematic of the experimental model used in ( B ) and ( C ). (B and C) Quantification of Ag-experienced CD4+CD45.2+ OT-II cells from lung and lung-draining lymph node after i.v. transfer into OVA-sensitized mice that were subsequently challenged i.t. with OVA in the presence or absence of anti-JAG1 blockade. *p ≤ 0.05 as determined by unpaired t test, n = 4–5 mice per group. ( D ) Diagram of the model used in ( E ) and ( F ). (E and F) Quantification of the number of CD4+CD45.2+ OT-II cells in the lung and lymph node of CD45.1 mice that received a transfer of naive CD45.2+ OT-II T cells and were subsequently challenged i.t. with OVA in the presence or absence of anti-JAG1 blockade. More
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