ccr4 inhibitor  (MedChemExpress)

Journal: Scientific Reports

Article Title: Ischemic stroke outcome after promoting CD4+CD25+ Treg cell migration through CCR4 overexpression in a tMCAO animal model

doi: 10.1038/s41598-024-60358-2

Figure Lengend Snippet: Chemokines observed in murine chemokine arrays in conditioned primary microglia culture media and interactions with known receptors. The numbers on the left (#) represent the chemokines shown in Fig. 1 d.

Article Snippet: The CCR4 inhibitor (sc-221406) was obtained from Santa Cruz Biotechnology, Inc. (Dallas, Texas), and the CXCR4 inhibitor (WZ811) was obtained from MedChemExpress (Monmouth Junction, New Jersey).

Techniques: Activation Assay, Migration, Cell Differentiation, Activity Assay

Journal: Scientific Reports

Article Title: Ischemic stroke outcome after promoting CD4+CD25+ Treg cell migration through CCR4 overexpression in a tMCAO animal model

doi: 10.1038/s41598-024-60358-2

Figure Lengend Snippet: Confirmation of the selected receptors in cytotoxic T cells and Treg cells through immunocytochemistry (ICC) and western blot. ( a ) ICC confocal imaging results showing the expression of the screened receptors: CXCR4, CD4, CCR4, and CCR10 on cytotoxic T cells and Treg cells, as well as CD8α or FoxP3, respectively. ( b ) The corrected total cell fluorescence (CTCF) was measured with ImageJ and normalized against either CD8α or FoxP3, respectively. ( c ) Western blot results of the receptors listed above, as well as β-actin as a reference in cytotoxic T cells and Treg cells. Each receptor from each cell type was ran in different gels and transferred onto separate membrane (full length gels can be viewed in the Supplementary Fig. ). ( d ) The above western blot band intensity for each receptor was measured using ImageJ and normalized against the β-actin bands in cytotoxic T cells and Treg cells. Asterisks denote the significance of data from p-values obtained by performing the Two-way ANOVA and Sidak’s multiple comparison (* p ≤ 0.05, ** p ≤ 0.01, **** p < 0.0001, all n = 3).

Article Snippet: The CCR4 inhibitor (sc-221406) was obtained from Santa Cruz Biotechnology, Inc. (Dallas, Texas), and the CXCR4 inhibitor (WZ811) was obtained from MedChemExpress (Monmouth Junction, New Jersey).

Techniques: Immunocytochemistry, Western Blot, Imaging, Expressing, Fluorescence, Membrane, Comparison

Journal: Scientific Reports

Article Title: Ischemic stroke outcome after promoting CD4+CD25+ Treg cell migration through CCR4 overexpression in a tMCAO animal model

doi: 10.1038/s41598-024-60358-2

Figure Lengend Snippet: The Treg cell MSCV-CCR4 retrovirus transduction showed an increase in CCR4 expression, further increasing the migration into the OGD media. ( a ) The MSCV retrovirus vector construct was designed via VectorBuilder. The mCcr4 sequence was linked with the EGFP reporter gene with a P2A linker, where the expression was controlled by the MSCV ψ+ promoter. ( b ) Transduction was performed following this schedule. ( c ) ICC imaging of the UT and TD Treg cells. ( d ) CTCF was measured with the ImageJ program and normalized against the FoxP3 fluorescence measurements. ( e ) The detailed quantification of changes in CCR4 expression was observed using FACS. ( f , g ) Graph of the FoxP3+ Treg cell population expressing EGFP and CCR4 highly. ( h ) Confocal images of Treg cells that migrated into the chamber containing OGD media. ( i ) The number of Treg cells that migrated into the OGD media after 48 h. Asterisks denote the significance of the p-values obtained by performing the unpaired t-test (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p < 0.0001, all n = 3–5).

Article Snippet: The CCR4 inhibitor (sc-221406) was obtained from Santa Cruz Biotechnology, Inc. (Dallas, Texas), and the CXCR4 inhibitor (WZ811) was obtained from MedChemExpress (Monmouth Junction, New Jersey).

Techniques: Transduction, Expressing, Migration, Plasmid Preparation, Construct, Sequencing, Imaging, Fluorescence

Journal: Neuro-oncology Advances

Article Title: Immune deconvolution and temporal mapping identifies stromal targets and developmental intervals for abrogating murine low-grade optic glioma formation

doi: 10.1093/noajnl/vdab194

Figure Lengend Snippet: CD8 + T cell predominance is governed by NF1/RAS regulation of Ccr4 expression. A, CD8 + T cells content in Nf1 -OPG optic nerves ( n = 10) is similar to controls ( n = 8) at 3, 4.5, 6, and 9 WOA, but increases by 12 WOA ( P < 0.0001). Nf1 +/– mice ( n = 6) have similar numbers of CD8 + T cells as controls from 3 to 12 WOA. B, CD4 + T cell content does not change in Nf1 -OPG optic nerves relative to controls at 3, 4.5, 6, 9, and 12 WOA. C, Ccl2 did not increase CD4 + T cell migration in WT or Nf1 +/– mice. Ccl12 or medium (control) does not increase CD4 + T cell migration. Ccl2 increased Nf1 +/– CD8 + T cell migration relative to WT controls ( P < 0.0001). Nf1 +/- and WT CD8 + T cells treated with medium (Control) or Ccl12 exhibit similar migration. D, Nf1 +/- and WT CD4 + T cells had similar levels of Ccr4 expression. Nf1 +/– CD8 + T cells have increased Ccr4 expression relative to their WT counterparts ( P = 0.0474). E, Ccr4 inhibition (AZD2098; 15µM) in CD8 + T cells decreases Ccl2-induced, but not Ccl12-induced, migration ( P = 0.0063). F, Nf1 +/– and WT CD4 + T cells had similar levels of Ras activity. Nf1 +/– CD8 + T cells have increased Ras activity relative to their WT counterparts ( P = 0.0363) G, Inhibition of Ras activity (10µM IN-1) reduces Ccl2-induced,but not Ccl12-induced, CD8 + T cell migration ( P = 0.0104). H, RAS activity inhibition (10µM IN-1) decreases Ccr4 expression in Nf1+/- CD8 + T cells ( P = 0.0069). I, Proposed model of Ccl2 signaling through Ccr4 in Nf1+/– CD8 + T cells.

Article Snippet: After 24h, Nf1 +/- CD8 + T cells were treated for 24 h with 10 μM Pan-RAS-Inhibitor-IN-1 (IN-1[HY-101295, MedChem Express]) or 10 μM Ccr4 inhibitor (AZD2098, Selleckchem).

Techniques: Expressing, Migration, Control, Inhibition, Activity Assay