lps (e. coli serotype 0111:b4  (Millipore)

Journal: bioRxiv

Article Title: Acute rapamycin treatment reveals novel mechanisms of behavioral, physiological, and functional dysfunction in a maternal inflammation mouse model of autism and sensory over-responsivity

doi: 10.1101/2024.07.08.602602

Figure Lengend Snippet: ( A ) Experimental timeline summary; ( B ) Inflammatory blood cytokines are significantly increased in both young and old adult MIR offspring relative to age matched controls (all p<0.001 MIR greater than control values) and are expressed as fold-change in MIR relative to controls (G-CSF=granulocyte colony stimulating factor, IFN-g=interferon gamma, IL- 6=interleukin 6, IP-10=interferon gamma induced protein, MCP-1=monocyte chemoattractant, MIG= monokine induced by gamma protein interferon, VEGF=vascular endothelial growth factor, KC=keratinocyte chemoattractant, M-CSF=macrophage colony stimulating factor, GM- CSF=granulocyte macrophage colony stimulating factor, IL-9=interleukin 9, IL-1b=interleukin 1 beta, TNF-a=tumor necrosis factor alpha, LIF=leukemia inhibitory factor; all cytokines; ( C ) Blood cytokine changes in LPS injected pregnant dams (0.008 mg/kg) graphed as fold-change relative to vehicle- injected control pregnant dams at 12 hours post-injection, all cytokines (p<0.01); ( D ) Stereoinvestigator quantification of IBA+ immunohistochemistry staining of microglia in the sensory- motor cortex of adult MIR offspring, graphed as % control, all *p<0.05 (MIR v control values); ( E ) Phospho-S6 protein measured by western blot on microdissected, flash-frozen brain tissue from cortex, amygdala, and striatum (combined),*p=0.022;); ( F ) Repetitive behaviors that are significantly increased in young and old MIR mice compared to controls (*p<0.01) are significantly reduced after 3 week Plexxikon 5622 treatment to ablate brain microglia in the young (**p<0.05) but not the old adult MIR offspring; ( G ) Social approach (ratio of time spent interacting with mouse relative to empty cup) in the 3 chamber social preference test is significantly reduced in young and old adult MIR mice (*p<0.01) but is significantly increased after 3 week Plexxikon 5622 treatment in young (**p<0.05) but not old MIR offspring; ( H ) Sensitivity to touch/pressure on their hind paws in the Von Frey fiber sensitivity test is significantly lower in young and old adult MIR mice (*p<0.05) but is only increased toward control levels in young MIR mice after 3 week Plexxikon 5622 treatment (**p<0.05); all data mean ± SEM

Article Snippet: Maternal Inflammatory Response (MIR) was induced with low-dose lipopolysaccharide (LPS, E. coli serotype 0111:B4; 0.008 mg/kg, Sigma) administered in a single I.P. injection on E9 of gestation as described in Le Belle, et al., 2014.

Techniques: Control, Injection, Immunohistochemistry, Staining, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: A small-molecule allele-selective transcriptional inhibitor of the MIF immune susceptibility locus

doi: 10.1016/j.jbc.2024.107443

Figure Lengend Snippet: Impact of CMFT on cellular MIF RNA and protein expression. A , human THP-1 monocytes transfected with MIF promoter-luciferase reporter plasmids bearing 0, 5, 6, 7, and 8 CATT repeats were stimulated with lipopolysaccharide (LPS, 100 ng/ml), treated with CMFT (3 μM) or vehicle control (0.4% DMSO) for 6 h and luciferase activity assessed by Dual-Luciferase assay. B , quantitative PCR analysis of MIF mRNA of bone marrow-derived macrophages (BMDMs) isolated from humanized MIF CATT5 and MIF CATT7 mice, stimulated in vitro with LPS (100 ng/ml), and treated with CMFT (2.5 μM, 6 h) or vehicle control (0.4% DMSO). C , ELISA analysis of human MIF in supernatants (triplicate measurements) of cultured LPS-stimulated (100 ng/ml) BMDMs from MIF CATT5 and MIF CATT7 mice 6 h after treatment with CMFT (2.5 μM) or vehicle control (0.4% DMSO). Data are the mean+SD and representative of two replicated experiments (∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 by Student’s t test, two-tailed).

Article Snippet: Transfected monocytes were stimulated with lipopolysaccharide (LPS, 100 ng/ml; E. coli serotype 0111:B4, Sigma-Aldrich) and simultaneously treated with test compounds or vehicle control (0.4% DMSO), and the luciferase activity assessed 6 h later by Dual-Luciferase assay (Promega).

Techniques: Expressing, Transfection, Luciferase, Control, Activity Assay, Real-time Polymerase Chain Reaction, Derivative Assay, Isolation, In Vitro, Enzyme-linked Immunosorbent Assay, Cell Culture, Two Tailed Test