Song, Ting-Ting Bi, Yan-Hua Gao, Yu-Qi Huang, Rui Hao, Ke Xu, Gang Tang, Jia-Wei Ma, Zhi-Qiang Kong, Fan-Ping Coote, John Chen, Xue-Qun Du, Ji-Zeng Additional file 1: Figures S1–S3. of Systemic pro-inflammatory response facilitates the development of cerebral edema during short hypoxia Figure S1. (A) Brain water content was measured following injection of rats with LPS (ip) for 12 h, H2O-injection for 0.5 h or both (n = 6). ***p < 0.001 compared with control; ### P < 0.001 compared between 20 % ddH2O and 20 % ddH2O + LPS treatment. (B) Immunohistochemical fluorescence of GFAP in the cortex of rats treated as indicated. Scale bar, 50 μm. Representative transmission electron micrographs of perivascular astrocytes and mitochondria. Scale bar, 1 μm (last one). Figure S2. (A) Fluorescence intensity of RFP-AQP4 and water permeability were measured in transfected RFP-AQP4 HeLa cells (n = 60). (B) Astrocytes were pre-treated with CHX or ActD or untreated. The water permeability of astrocytes was determined after 12 h LPS stimulation (n = 100). ***P < 0.001 compared with control; ### P < 0.001 compared with LPS 12 h. (C) Brain water content was determined by dry-wet weight method at the indicated time points and under the indicated conditions (n = 6–12). ***p < 0.001 compared with control; ### P < 0.001 compared with LPS + hypoxia, all data are presented as mean ±SD. Figure S3. Inflammation induced AQP4 high expression through MAPK pathway. (A) Astrocytes were treated with LPS as indicated time, and the phosphorylation levels of ERKs and p38 were tested by western blot using anti-phospho-ERK or anti-phospho-p38 antibodies (n = 3). *P < 0.05; **P < 0.01;***P < 0.001 compared with control. (B) Acetylation levels of H3 and H4 in astrocytes were determined by western blot as indicated time after LPS treatment (n = 3). ***P < 0.001 compared with control, Student’s t test. (C,D) Increased-AQP mRNA by LPS in astrocytes were be blocked by Cur, si-p300. ***P < 0.001 compared with control, ### P < 0.001 compared with LPS or si control, all data are presented as mean ±SD. (DOC 1687 kb) DOC 1687  kb;CHX;2O;transfected RFP-AQP 4 HeLa cells;LPS 12  h;Brain water content;LPS treatment;anti-phospho-p 38 antibodies;dry-wet weight method;water permeability;astrocyte;SD;1 μ m;scale bar;hypoxia Figure S 1.;Systemic pro-inflammatory response;12  h LPS stimulation;Figure S 2.;Figure S 3. Inflammation;representative transmission electron micrographs;50 μ m;ERK;GFAP;MAPK 2016-03-11
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