4/6/2023 0 Comments Mtmr 13 antibody![]() ![]() (D) HE staining, ZO-1, Occludin and Ki67 immunofluorescent staining in Rag1 -/- mice, scale bar is 100 mum (n = 8). (A-C) Intestinal lamina propria cells were analyzed by flow cytometry for the ratio of ILC2/ILCs and IL-13 + ILC2/ILC2 in Rag1 -/- mice (n = 3-4). ( K-L ) Tumor of the mouse models from ( F ) were evaluated for infiltration of both CD4 + and CD8 + T cells on day 16 post implantation, whichįigure 5 Depletion of ILC2 abolished the protective effect of IL-33 on intestinal I/R injury and organoids H/R injury. ( I-J ) 6PGD blockade in Tregs resulted in lower tumor volume ( I ) and tumor weight ( J ). ( H ) Suppression assay on sorted EGFP + Tregs as described in supplemental information. ( G ) Flow cytometry analysis show the same frequency of Tregs (EGFP + ) in CD4 + cells in spleen of both mouse groups 10 days post tamoxifen treatment. ( F ) Tamoxifen treatment schedule and tumor induction by implanting B16F10 cells in Pgd fl/fl Foxp3 EGFP-Cre-ERT2 and WT mice. YFP + cells were sorted for the suppression assay. ( D-E ) 6-AN treatment of driven Tregs demonstrate lower suppressive capacity evaluated in in vitro suppression assay. ( C ) Real-time PCR analysis on derived Tregs demonstrate lower expression of Foxp3 transcription factor and higher expression of Gata3 transcription factor and IL-5 under treatment with 6-AN versus vehicle DMSO. YFP + percentage in CD4 + T cells ( A ) and YFP gMFI ( B ) are demonstrated. ( A-B ) Wild-type (WT) Foxp3 Cre naive CD4 + T cells (CD62L high CD44 low ) were driven toward Tregs in vitro (as inducible Treg ) along with treatment with 6-aminonicotinamide (6-AN) or vehicle DMSO and Treg drive efficacy was evaluated as CD25 + YFP + cells. 6-Phosphogluconate dehydrogenase (6PGD) blockade in regulatory T cells (Tregs) prevents their suppressive function and induces potent anti-tumor responses. (G) Representative plots and quantitative graphs of the freįigure 3. Cells were gated on live CD45 + CD19 - TCRbeta + CD4 + (data pooled from 2 independent assays). (F) Representative plots and quantitative graphs of the frequency and numbers of 2W1S-specific CD4 + T cells in lung parenchyma from H2-Ab1 fl/fl control mice and MHC class II DeltaILC3 mice after exposure to papain plus 2W1S peptide or PBS control. (D and E) Representative plots of MHC class II expression in designated immune cells (D) and quantification of MHC class II staining on those cell types in the MedLN following HDM exposure (E) (representative of 2 independent assays, n = 4). Naive T cells were gated as CD44 - CD62L +, effector T cells were gated as CD44 + CD62L -, and Tregs were gated as FoxP3 +. (C) Graphs quantifying CD25 staining or IL-2 binding in gated MedLN cells following HDM exposure (pooled from 2 independent assays, n = 10). (B) Histograms demonstrating CD80 and CD86 expression in MedLN cells following HDM exposure (representative of 2 independent assays containing 8 mice each). Dashed line indicates the B cell follicle border. Images taken at 20x magnification scale bars, 50 mum (left panel) and 10 mum (inserts right panel). ILC3s limit CD4 + T cell responses to allergens in the airway (A) Representative immunofluorescence staining for ILC3s and T cells in the MedLNs of HDM-exposed mice (representative of 2 independent experiments and n = 3 mice per group). PA, pravastatin I/R, ischemia/reperfusion ILC2, type II innate lymphoid cells.įigure 2. * p < 0.05, ** p < 0.01, *** p < 0.001, NS means No statistically significant difference by one-way ANOVA (Tukey's test). The results are expressed as the mean +- SEM (B, C, E-J, N). (M, N) IL-13 immunohistochemical staining and intensity quantification analysis in the ileum, scale bar is 100 mum (n = 8). (K, L) Intestinal lamina propria cells were analyzed by flow cytometry for the ratio of ILC2/ILCs and IL-13 + ILC2/ILC2 in WT mice (n = 3-4). (J) The relative IL-33 intensity quantification analysis in the ileum. ![]() (I) The relative mRNA level of Lgr5 in the ileum was measured by quantitative PCR (n = 8). (F-H) The relative fluorescence intensity quantification analysis of ZO-1, Occludin and Ki67 in the ileum. (E) Pathological damage score in the ileum. (D) HE staining, ZO-1, Occludin and Ki67 immunofluorescent staining and IL-33 immunohistochemical staining in WT mice and IL-33 -/- mice, scale bar is 100 mum (n = 8). (B, C) Relative mRNA levels of IL-33 and IL-33 receptor (ST2) (n = 8). (A) IL-33 immunohistochemical staining in the ileum from sham, I/R and I/R + PA mice, scale bar is 100 mum (n = 8). Figure 3 PA protected against intestinal I/R injury via an IL-33/ST2 signal. ![]()
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