Additional file 1 of Brain profiling in murine colitis and human epilepsy reveals neutrophils and TNFα as mediators of neuronal hyperexcitability
Creators: name: Barnes, Sarah E.
name: Zera, Kristy A.
name: Ivison, Geoffrey T.
name: Buckwalter, Marion S.
name: Engleman, Edgar G.
Publisher: name: figshare
Year: 2021
Subjects: subject: Biophysics
subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Neuroscience
subject: Physiology
subject: Pharmacology
subject: Immunology
subject: FOS: Clinical medicine
subject: Marine Biology
subject: 111714 Mental Health
subject: FOS: Health sciences

description: Additional file 1: Supplemental Figure 1. Linear regressions between cytokine transcription and Tjp1 and Cldn5. n = 5-10 mice/group, representative of 3-5 replicate experiments. Supplemental Figure 2. Frequencies of immune cell populations in the brains of mice following acute DSS-induced colitis. Flow cytometric quantification of a) macrophages (CD45+CD11b+Ly6G-Ly6C-MHCII+), b) dendritic cells (CD45+CD11b+Ly6G-CD11c+), c) B cells (CD45+CD11b-CD3-CD19+), and d) T cells (CD45+CD11b-CD3+). n = 5-10 mice/group, representative of 3-5 replicate experiments. ns=not significant, *p &lt; 0.05. Supplemental Figure 3. Depletion efficiency of monocytes and neutrophils in mice following acute DSS-induced colitis. a-b) 200mL of clodronate-loaded liposomes was administered on days 6-14 during acute DSS-induced colitis. a) Flow cytometric quantification of monocytes (CD45+CD11b+Ly6G-Ly6C+MHCII-) in colitic mouse blood following clodronate treatment. b) Percent of monocytes in colitic mouse blood following clodronate treatment normalized to the mean percent of monocytes in colitic mice treated with PBS-loaded liposomes. c-d) 500mg α-Ly6G antibody was administered on days 6-14 during acute DSS-induced colitis induction. c) Flow cytometric quantification of neutrophils (CD45+CD11b+Ly6G+) in colitic mouse blood following α-Ly6G antibody treatment. d) Percent of neutrophils in colitic mouse blood following α-Ly6G antibody treatment normalized to the mean percent of neutrophils in colitic mice treated with isotype IgG2a antibody. Data are representative of 3-4 combined experiments with 5-15 mice/group/experiment. *p &lt; 0.05, ****p &lt; 0.0001. Supplemental Figure 4. Representative images of in vivo α-CD45 staining of border-associated immune cells. a) Immunofluorescence images of the choroid plexus in healthy and colitic mice following in vivo staining of border regions with an α-CD45 antibody and subsequent ex vivo α-CD45 staining of all leukocytes. Parenchymal cells (i.e. microglia) do not stain positive for the in vivo-administered α-CD45, but border-associated cells (i.e. those in the choroid plexus) stain for both the in vivo- and ex vivo-administered α-CD45. b) Representative flow cytometry plots of in vivo- and ex vivo-administered α-CD45 staining of microglia (CD45loCX3CR1hiCD11b+) and neutrophils (CD45+CD11b+Ly6G+) in the brains of healthy and colitic mice. Microglia (parenchymal cells) are not stained by the in vivo-administered α-CD45 in either condition. 031821 J Neuroinflammation Tables.pdf. Supplementary Tables.