NaCl / 125 / 7.305 / 73.05
KCl / 2.5 / 0.186 / 1.86
MgCl.6H2O / 1 / 0.2033 / 2.033
NaH2PO4 / 1.25 / 0.15 / 1.5
CaCl2.2H2O / 2 / 0.294 / 2.94
NaHCO3 / 25 / 2.1025 / 21.025
Glucose / 25 / 4.505 / 45.05
pH with NaOH to 7.3
TableS1: Composition of artificial cerebrospinal fluid
Cells/Proteins of Interest / Primary Antibody / Dilution(In Normal Horse Serum) / Secondary Antibody / Retrieval Buffer
Microglia / IBA1 / 1:1000 / Anti-Goat / Citrate
Astrocytes / GFAP / 1:40,000 / Anti-Rabbit / Citrate
Table S2: Antibodies employed within this study
Legend: IBA1: Microglial cell marker, GFAP: Astrocytic cell marker for glial fibrillary acidic protein
Table S3: List of behavioural tests with P value and Mean ± SEM for LPS-aCSF mice compared to Sal-aCSF mice
Behavioural Test / Mean ± SEM(LPS-aCSF vs saline-aCSF) / Unpaired t-test
(LPS-aCSF vs Saline-aCSF)
OFT- Distance Travelled / 13.8 ± 0.73 vs.10.7 ± 0.79 / P = 0.01
EZM –Time in Open Arm / 45.8 ± 6.3 vs. 26.3 ± 2.3 / P = 0.031
EZM- Head Dip / 9.45 ± 1.9 vs 0.93 ± 0.3 / P = 0.0003
EZM- Stretch/Attend / 12.5 ± 2.4 vs 1.77 ± 0.5 / P = 0.0003
Y- Maze- % novel arm time / 67.6 ± 3.8 vs 53.6 ± 1.2 / P = 0.0048
IBA1 expression / 6.88 ± 0.6 vs 14.06 ± 0.8 / P = 0.0002
GFAP expression / 13.5 ± 1.3 vs 12.2 ± 0.4 / P = 0.4060
Legend: The table lists the Mean ± SEM and P value for LPS-aCSF group compared to Saline-aCSF in the Open Field Test (OFT), Elevated Zero Maze (EZM) and Y maze. The table also shows the Mean ± SEM and P value for number of microglial (IBA1) and astrocytic (GFAP) positive cells in the dentate gyrus.
Table S4: List of behavioural tests with Mean ± SEM and P value for LPS-Etan group compared to LPS-aCSF
Behavioural Test / Mean ± SEM (LPS-Etan vs LPS-aCSF) / Unpaired t-test (LPS-Etan vs LPS-aCSF)OFT –Distance Travelled / 9.84 ± 0.5 vs. 10.7 ± 0.7 / P = 0.37
EZM – Open arm time / 56.7 ± 9.5 vs. 26.3 ± 2.3 / P = 0.03
EZM- Head Dip / 7.11 ± 1.7 vs. 0.93 ± 0.3 / P = 0.0039
EZM- Stretch/Attend / 12.3 ± 2.4 vs.1.78 ± 0.5 / P = 0.0008
Y- Maze- % novel arm time / 62.1 ± 6.5 vs. 53.6 ± 1.2 / P = 0.1249
IBA1 expression / 10.4 ± 0.4 vs. 14 ± 0.8 / P = 0.0127
GFAP expression / 16.8 ± 2.4 vs. 12.2 ± 0.4 / P = 0.07
Legend: The table lists the Mean ± SEM and P value for LPS-Etan group compared to LPS-aCSF in the Open Field Test (OFT), Elevated Zero Maze (EZM) and Y maze. The table also shows the Mean ± SEM and P value for number of microglial (IBA1) and astrocytic (GFAP) positive cells in the dentate gyrus.
Figure Legends
Figure S1:Astrocyte counts across experimental conditions
Legend:A) The figure shows IHC images of the astrocytes in the dentate gyrus while the graph represents the number of GFAP positive cells in the dentate gyrus. B) The figure shows IHC images of the astrocytesin the PFC while the graph represents the number of GFAP positive cells in the PFC. All data represent mean ± SEM (n = 5/group, Sal-aCSF: Saline treatment IP with aCSF administered ICV, Sal-Etan: Saline treatment IP with etanercept administered ICV, LPS-aCSF: LPS challenge IP with aCSF administered ICV, LPS-Etan: LPS challenge IP with etanercept administered ICV). Data compared using 2-way ANOVA (A: Interaction: F(1,15)=1.24, P=0.28, Saline/LPS challenge: F(1,15)=0.077, P=0.78,aCSF/etanercept treatment: F(1,15)=3.58, P=0.07, Fig S1A); (Interaction: F(1,12)=0.11, P=0.73, Saline/LPS challenge: F(1,12)=0.69, P=0.42aCSF/etanercept treatment: F(1,12)=0.63, P=0.44, Fig S1B).