Journal: Cell host & microbe

Article Title: Commensal Microbes and Hair Follicle Morphogenesis Coordinately Drive Treg Migration into Neonatal Skin

doi: 10.1016/j.chom.2017.03.001

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: K5-rtTA x tetO-Dkk1 mice (mixed genetic background) , Provided by Sarah Millar; Chu et al., 2004 ; Kistner et al., 1996 , N/A.

Techniques: Virus, Plasmid Preparation, Recombinant, cDNA Synthesis, SYBR Green Assay, Cell Isolation, Selection, RNAscope, Staining, RNA Sequencing, Software

Journal: Journal of Neuroinflammation

Article Title: EBI2 receptor regulates myelin development and inhibits LPC-induced demyelination

doi: 10.1186/s12974-017-1025-0

Figure Lengend Snippet: EBI2 deficiency results in transient delay in MBP expression. a Representative confocal images show mice cerebellar slices prepared from P10 mice and cultured for 1, 2, 4 to 5, 7 to 9, and 14 DIV. Myelin (MBP, green) staining is shown. Myelin development was determined by quantification of MBP staining density. Analysis showed significant differences in myelin development between one to two DIV. Images taken at ×10 magnification. Scale bar, 100 μM. b Graph showing quantification of MBP immunofluorescence in images such as those shown in ( a ). Data presented as mean ± SD (five to six animals per group), unpaired t test, ** p < 0.01, *** p < 0.001 versus corresponding control. c Representative WBs show differential expression of MBP in whole brain lysates prepared from embryonic (E14 and E18) and postnatal (P1, P7, P14, and P30) WT and EBI2 KO mice. d, e Densitometric quantification of blots such as that shown in ( c ). Normalized WB data presented as mean ± SD (three to four WB data per time point; one animal per blot), unpaired t test, *** p < 0.001 versus corresponding control

Article Snippet: The EBI2 double (−/−) KO mouse strain (mixed genetic background C57BL/6 x C129) was purchased from Deltagen [ ].

Techniques: Expressing, Cell Culture, Staining, Immunofluorescence, Control, Quantitative Proteomics

Journal: Journal of Neuroinflammation

Article Title: EBI2 receptor regulates myelin development and inhibits LPC-induced demyelination

doi: 10.1186/s12974-017-1025-0

Figure Lengend Snippet: Downregulation or antagonism of EBI2 signaling staggers myelination. Representative confocal images show MBP expression (green) and NF light chain (red) in WT cerebellar slices prepared from P10 mice and cultured for a 9 DIV, b 14 DIV, and c 20 DIV with or without 7α25HC (1 μM) and NIBR189 (1 μM). Corresponding bar graphs shows quantification of MBP staining. There are no differences between non-treated and treated slices at 9 and 14 DIV. However, treatment of slices with 7α25HC or NIBR189 for 20 DIV inhibits normal myelin development in WT cerebellar slices as measured by MBP staining. Data presented as mean ± SD ( n = 4 and 6), one-way ANOVA and Dunnett’s post-test, ** p > 0.01, *** p < 0.001 versus corresponding control. Images taken at ×10 magnification. Scale bar 100 μm ( d ). Treatment of human monocytes (U937) with 7α25HC (10 μM) for 1 h induces EBI2 (green) internalization. NIBR189 (10 μM) does not induce receptor internalization. Images were taken at ×40 magnification. Scale bar 20 μM

Article Snippet: The EBI2 double (−/−) KO mouse strain (mixed genetic background C57BL/6 x C129) was purchased from Deltagen [ ].

Techniques: Expressing, Cell Culture, Staining, Control

Journal: Journal of Neuroinflammation

Article Title: EBI2 receptor regulates myelin development and inhibits LPC-induced demyelination

doi: 10.1186/s12974-017-1025-0

Figure Lengend Snippet: 7α25HC protects from LPC-induced demyelination. Representative confocal images show WT ( a ) and EBI2 KO ( b ) mice cerebellar slices treated with 7α25HC (1 μM) and/or NIBR189 (1 μM) ± LPC (0.4 mg/ml). Anti-MBP (green) and anti-neurofilament (NF, red) immunostaining is shown. Images were taken at ×10 magnification. Scale bar, 100 μM. c A corresponding bar graph shows quantification of myelin staining in WT slices such as those shown in ( a ). 7α25HC significantly protects from LPC-induced demyelination, and this protective effect is inhibited with the EBI2 antagonist NIBR189 indicating EBI2 mediated protection from demyelination. Data presented as mean ± SD ( n = 4 and 8), one-way ANOVA and Tukey’s multiple comparison post-test, ** p < 0.01 versus corresponding control. d A bar graph shows that 7α25HC does not protect from LPC-induced demyelination in slices prepared from EBI2 KO animals confirming EBI2-dependent mechanism shown in ( a and c ). Data presented as mean ± SD ( n = 4), one-way ANOVA and Tukey’s multiple comparison post-test, ** p < 0.01 versus control. e Schematic diagram illustrating experimental design

Article Snippet: The EBI2 double (−/−) KO mouse strain (mixed genetic background C57BL/6 x C129) was purchased from Deltagen [ ].

Techniques: Immunostaining, Staining, Comparison, Control

Journal: Journal of Neuroinflammation

Article Title: EBI2 receptor regulates myelin development and inhibits LPC-induced demyelination

doi: 10.1186/s12974-017-1025-0

Figure Lengend Snippet: 7α25HC attenuates LPC-induced levels of IL1β and IL6 in cerebellar slices. a Schematic diagram illustrating experimental design. Treatment of slices with LPC (0.4 mg/ml) induces an increase in IL6 ( b ) and IL1β ( c ) protein release. 7α25HC (1 μM) attenuates LPC (0.4 mg/ml)-induced levels of IL6 ( b ) and IL1β ( c ). Co-treatment with EBI2 antagonist (NIBR189, 1 μM) blocks the 7α25HC-mediated effects on the levels of IL6 ( b ) and IL1β ( c ) indicating EBI2-dependent regulation of cytokine release. Treatment with NIBR189 alone did not alter IL6 ( b ) or IL1β ( c ) release. Data presented as mean ± SD ( n = 3), one-way ANOVA and Bonferroni’s post-test, *** p < 0.001 versus corresponding control

Article Snippet: The EBI2 double (−/−) KO mouse strain (mixed genetic background C57BL/6 x C129) was purchased from Deltagen [ ].

Techniques: Control