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Black dotted line denotes no change in relative abundance of transcripts.
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Results normalized to β2m mRNA levels and relative to vehicle-treated controls n = 5–9 mice/treatment group, representing 2–3 independent experiments. (E) Relative transcript abundance of IFN-γ and Tbx21 in the labiar tissue at 1 and 21 days after 10 th MI challenge. Results with no detectable cells displayed as “n.d.” Significance with respect to control groups * p<0.5, ** p<0.01, *** p<0.001. Results displayed as mean ± SEM, outliers excluded via Grubbs testing n = 3-6/treatment group, 1 sample representing 2 mice pooled.
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Results displayed as mean ± SEM, outliers excluded via Grubbs testing n = 3-6/treatment group, 1 sample represents 2 mice pooled representing 2 independent experiments. Total counts were calculated using beads and FlowJo gating counts and percentages, cell counts for each marker were calculated with their respective frequency of live cells and total cell count.
#HOW TO CHANGE DASHED LINE IN FLOWJO 10 SKIN#
(C-D) Flow cytometric based detection of CD8 + cells in flank skin collected 1 day or 21 days after 10 th challenge (C) or 1 day after 1 st or 3 rd challenge (D). (A-B) Flow cytometric analysis of collagenase-digested CD4 + cells, gradient separated from MI- or AOO-treated flank skin collected at 1 and 21 days after the cessation of 10 challenges (A) or 1 day after 1 st or 3 rd challenge (B). Significance with respect to control group * p<0.05.ĬD4 + and CD8 + T cell infiltration, alongside upregulation of IFN-γ and Tbx21 mRNA transcripts, is induced by repeated MI challenge in previously sensitized mice. (G) Tissue myeloperoxidase levels measured as optical density/g wet labiar tissue at 1 day after 10 MI challenges n = 5-7/treatment group, representative of 2 separate experiments. Significance with respect to control group ** p<0.01. (F) Tissue eosinophil peroxidase levels measured by optical density/g wet labiar tissue 1 day and 21 days after 10 th MI challenge n = 2-7/treatment group, representative of 2 separate experiments. Significance with respect to control group ** p<0.01, *** p<0.001. (C-E) Protein concentration of Cxcl2 (C), IL-1β (D) and IL-6 (E) 1 and 21 days after 10 MI labiar challenges, determined via ELISA and normalized to total protein concentration from DC assay n = 3-8/treatment group. Results normalized to housekeeping gene β2m mRNA levels and vehicle challenged controls n = 6/treatment group, representative of 2 independent experiments. (B) Relative transcript abundance of Cxcl2, IL-1β, and IL-6 21 days after 10 daily MI challenges. Results normalized to housekeeping gene β2m mRNA levels n = 9/treatment group, representative of 3 separate experiments. (A) Transcript abundance of Cxcl2, IL-1β, and IL-6 in labiar tissue extracted 1 day after 10 daily challenges on the labia relative to vehicle challenged controls. Repeated MI challenge induces upregulated gene expression and protein content of pro-inflammatory cytokines, and increased eosinophil and neutrophil activity in the labiar skin of outbred ND4 Swiss mice.