Additional file 3 of Diesel exhaust impairs TREM2 to dysregulate neuroinflammation
Creators: name: Greve, Hendrik J.
name: Mumaw, Christen L.
name: Messenger, Evan J.
name: Prasada R. S. Kodavanti
name: Royland, Joyce L.
name: Kodavanti, Urmila P.
name: Block, Michelle L.
Publisher: name: figshare
Year: 2020
Subjects: subject: Biophysics
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Neuroscience
subject: Physiology
subject: Ecology
subject: Immunology
subject: FOS: Clinical medicine
subject: FOS: Clinical medicine
subject: Marine Biology
subject: 110309 Infectious Diseases
subject: FOS: Health sciences
subject: FOS: Health sciences
subject: Computational Biology

description: Additional file 3: Figure S1. Diesel Exhaust Exposure Fails to Affect Markers of Antioxidant System. WKY rats were exposed to diesel exhaust (0, 50, 150, or 500 μg/m3) by inhalation for 4 weeks. Key markers of the endogenous antioxidant system, Nfel2l (NRF2) (A) and Hmox1 (Heme Oxygenase 1) (B) were analyzed by qRT-PCR in the cortex. Data are reported as the mean ± SEM. (n = 7). Figure S2. Diesel Exhaust Exposure Fails to Affect Markers of Blood Brain Barrier Function. WKY rats were exposed to diesel exhaust (0, 50, 150, or 500 μg/m3) by inhalation for 4 weeks. Key markers of the BBB, Aqp4 (Aquaporin-4), Mdr1b (p-Glycoprotein), Icam1 (ICAM-1), and Vcam1 (VCAM-1) were analyzed by qRT-PCR in the hippocampus. Data are reported as the mean ± SEM. (n = 7). Figure S3. Peripherally Administered LPS Increases Microglial Association with the Vasculature at 3 Hours. C57Bl/6 J mice were administered LPS (5 mg/kg, IP) or saline (IP). The changes in the association of microglia cell bodies with the neurovasculature were assessed with confocal images of microglia (IBA-1, green) and vascular endothelial cells (CD31, red) in the hippocampus (3 slices per brain). Representative maximum intensity projection images are shown (A). Quantification of the number of vessel-associated microglia in CA1 hippocampus (B). *p &lt; 0.05 when compared to control. (n = 3). Figure S4. TREM2 Deficiency Does Not Impact Neuroinflammatory Response to Peripherally Administered LPS at 3 Hours. C57Bl/6 J mice were administered LPS (5 mg/kg, IP) or saline (IP). Expression of the pro-inflammatory factors Tnf (A), Il1b (B), and the cytosolic PHOX component Ncf1 (C) were analyzed by qRT-PCR in the cortex. Data are reported as the mean ± SEM. *p &lt; 0.05 when compared to Trem2+/+ control. #p &lt; 0.05 compared between Trem2-/- control. (n = 5-6). Figure S5. Intratracheal DEP Fails to Affect Microglia-Vessel Association in the Cortex. Trem2-/- and Trem2+/+ mice were treated with DEP (100 μg/mouse IT, 2x per week, 4 weeks) or vehicle (IT, 2x per week, 4 weeks). The changes in the association of microglia cell bodies with the neurovasculature were assessed with confocal images of microglia (IBA-1, green) and vascular endothelial cells (CD31, red) in the cortex (4 slices per brain). Representative maximum intensity projection images are shown (A). Quantification of the number of vessel-associated microglia in layer V cortex (B). (n = 3-6). Figure S6. Intratracheal DEP Fails to Affect Microglia-Vessel Association in the Hippocampus. Trem2-/- and Trem2+/+ mice were treated with DEP (100 μg/mouse IT, 2x per week, 4 weeks) or vehicle (IT, 2x per week, 4 weeks). The changes in the association of microglia cell bodies with the neurovasculature were assessed with confocal images of microglia (IBA-1, green) and vascular endothelial cells (CD31, red) in the hippocampus (4 slices per brain). Representative maximum intensity projection images are shown (A). Quantification of the number of vessel-associated microglia in CA1 hippocampus (B). (n = 3-6). Figure S7. Trem2 mRNA is Expressed by Isolated Microglia and Immune Cells in Bronchoalveolar Lavage. C57Bl/6 J mice were treated with DEP (600 μg/mouse IT, single dose) or vehicle (50 μL IT). mRNA levels of Trem2 were measured by qRT-PCR in isolated microglial cells (A) and immune cells collected by bronchoalveolar lavage (B). Data are reported as the mean ± SEM. (n = 3-5). Figure S8. gp91PHOX Fails to Regulate DEP-Induced Neuroinflammation. PHOX-/- or PHOX+/+ control mice were treated with DEP (600 μg/mouse IT, single dose) or vehicle (50 μL IT). mRNA levels were measured by qRT-PCR for Tnf (TNFα) (A,B) and Trem2 (C,D) in the hippocampus (A,C) and cortex (B,D). Data are reported as the mean ± SEM. *p &lt; 0.05, compared with controls. (n = 3-6).