Additional file 1: of A novel role for protein tyrosine phosphatase 1B as a positive regulator of neuroinflammation

This contains Figures S1 and S2. Figure S1. The PTP1B inhibitor, CinnGel, suppressed LPS-induced NO production in microglial cells. BV-2 microglial cells were treated with LPS (100 ng/ml) for 24 h after 1 h pretreatment with the indicated concentrations of CinnGel. The nitrite content was measured using the Griess reaction (a) and cytotoxicity of PTP1Bi was assessed by the MTT assay (b). The data were expressed as the mean ± SEM (n = 3). *p < 0.05 versus LPS only, one-way ANOVA with Tukey’s multiple comparison test. Figure S2. The Src inhibitor, PP2, suppressed LPS-induced NO production in microglial cells. BV-2 microglial cells were treated with LPS (100 ng/ml) for 24 h after 1 h pretreatment with the indicated concentrations of PP2. The nitrite content was measured using the Griess reaction (a) and cytotoxicity of PTP1Bi was assessed by the MTT assay (b). The data were expressed as the mean ± SEM (n = 3). *p < 0.05 versus LPS only, one-way ANOVA with Tukey’s multiple comparison test. (PDF 174 kb

Chronic Sleep Deprivation-Induced Proteome Changes in Astrocytes of the Rat Hypothalamus

Sleep deprivation (SD) can influence cognition, memory, and sleep/wake homeostasis and can cause impairments in many physiological processes. Because the homeostatic control of the sleep/wake cycle is closely associated with the hypothalamus, the current study was undertaken to examine proteomic changes occurring in hypothalamic astrocytes following chronic partial SD. After chronic partial SD for 7 days, astrocytes were prepared from rat hypothalamus using a Percoll gradient method, and their proteome profiles were determined by LC–MS/MS. Comparisons of the proteome profiles of hypothalamic astrocytes revealed that chronic partial SD increased (≥1.5-fold) 89 proteins and decreased (≤0.7-fold) 50 proteins; these changes in protein expression were validated by western blot or immunohistochemistry. DAVID and IPA analyses of these proteins suggested that SD may influence gliotransmission and astrocyte activation. PPP2R1A, RTN4, VAMP-2, LGI-1, and SLC17A7 were identified and validated as the main targets of SD in astrocytes. Our results suggest that SD may modulate gliotransmission in the hypothalamus, thereby disturbing sleep/wake homeostasis and increasing susceptibility to neurological disease; however, further studies are required to confirm whether the proteome changes are specific to SD