Journal: Journal of Neuroinflammation

Article Title: Chi3l3: a potential key orchestrator of eosinophil recruitment in meningitis induced by Angiostrongylus cantonensis

doi: 10.1186/s12974-018-1071-2

Figure Lengend Snippet: Elevation of Chi3l3 expression in the brain is an important characteristic in non-permissive host mice but not in permissive host rats during AC infection. a Expression profile of significant TCGs after AC infection. The horizontal axis represents genes, and vertical coordinates represents time points. D0 is the normal control group. D2, D7, D14, and D21 represent 2, 7, 14, and 21 dpi, respectively. b Expression patterns of TCGs. The x -axis represents time (with the unit as day), and the y -axis represents the normalized gene expression levels. c The mRNA levels of Chi3l3 in brains of mice infected with AC at 0, 1, 3, 7, 14, and 21 dpi. Mouse β-actin mRNA was used as an internal control. * P < 0.05. infected groups vs control group; ## P < 0.01, 14 dpi group vs 21 dpi group. d The mRNA levels of Chi3l3 in brains of rats infected with AC at 0, 1, 3, 7, 14, and 21 dpi. Rat B2m and Hprt1 mRNA levels were used as internal controls. * P < 0.05, ** P < 0.01, infected groups vs control group. e The protein levels of Chi3l3 in brains of mice infected with AC at 0, 7, 14, 21, and 28 dpi. β-Actin was used as an internal control. * P < 0.05, ** P < 0.01, *** P < 0.001, infected groups vs uninfected groups in the corresponding part. f A gene set enrichment analysis of transcriptome data was performed by comparing the mRNA levels of CLPs and chitinases in brains of mice infected with AC at 0, 2, 7, 14, and 21 dpi. Data information: In c – e , data are presented as mean ± SD. (Student’s t test)

Article Snippet: The following primary antibodies were used: mouse JMJD3 (ab85392, Abcam), CREB1 (A11989, ABclonal), CEBPB (A0711, ABclonal), KLF4 (A11853, ABclonal), Y641 phosphorylated-STAT6 (ab54461, Abcam), STAT6 (5397, Cell Signaling Technology), Chi3l3 (#60130, STEMCELL Technology), PPARγ (81B8, Cell Signaling Technology), and β-actin (12262, Cell Signaling Technology).

Techniques: Expressing, Infection

Journal: Journal of Neuroinflammation

Article Title: Chi3l3: a potential key orchestrator of eosinophil recruitment in meningitis induced by Angiostrongylus cantonensis

doi: 10.1186/s12974-018-1071-2

Figure Lengend Snippet: Eosinophil percentage coordinates with Chi3l3 derived from inflammatory macrophages in brains of AC-infected mice. a Chi3l3 + cells were observed in a gate of SSC and Chi3l3 + (A), within the gate (A). CD45 hi F4/80 + and F4/80 + CD11b + populations are shown in (C) and (D), respectively. b Chi3l3 + cells were characterized by CCR2 + gate (C) or CX3CR1 + gate (F) separately. c (A–D) The co-localization of Chi3l3 or Iba1 (A), CD11b or Chi3l3 (B), and DAPI (C) in cerebrum of AC-infected mice. (D) is the merge of (A), (B), and (C). (E–H) Chi3l3 or Iba1 (E), CD11b or Chi3l3 (F), and DAPI (G) in the cerebellum of AC-infected mice. (H) is the merge of (E), (F), and (G). Scale bars indicate 20 μm. Scale bar in (I) indicates 5 μm. d The percentages of Chi3l3 + CD45 lo F4/80 + cells in BMNCs of mice at 7, 14, and 21 dpi are presented. *** P < 0.001, **** P < 0.0001, AC-infected groups vs control group. Chi3l3 + CD45 hi F4/80 pos cells in BMNCs of mice at 7, 14, and 21 dpi are presented. #### P < 0.0001, AC-infected groups vs control group. $$ P < 0.01, $$$ P < 0.001, $$$$ P < 0.0001. e The percentages of eosinophils in BMNCs of mice infected with AC at 7, 14, and 21 dpi. There were 5–10 mice per group. Ctrl indicates the concurrent control. * P < 0.05, **** P < 0.0001, AC-infected groups vs control group. ### P < 0.001, AC-infected 14 dpi group vs AC-infected 7 dpi group. $$ P < 0.01, AC-infected 21 dpi group vs AC-infected 14 dpi group. f The percentages of eosinophils in peripheral blood mononuclear cells (PBMCs) of mice infected with AC at 7, 14, and 21 dpi; 5–10 mice per group. Ctrl indicates the concurrent control. * P < 0.05, AC-infected groups vs control group. g Clophosome and saline were injected into BALB/c mice intravenously 1 day before of AC infection, followed by five intravenous challenges every 4 days. At 22 dpi, the brain samples were collected. h Brain Chi3l3 mRNA levels ( g ). i Histopathological changes of the brain ( g ) (scale bar, 50 μm). Data information: In ( d-h ), data are presented as mean ± SD. (Student’s t test)

Article Snippet: The following primary antibodies were used: mouse JMJD3 (ab85392, Abcam), CREB1 (A11989, ABclonal), CEBPB (A0711, ABclonal), KLF4 (A11853, ABclonal), Y641 phosphorylated-STAT6 (ab54461, Abcam), STAT6 (5397, Cell Signaling Technology), Chi3l3 (#60130, STEMCELL Technology), PPARγ (81B8, Cell Signaling Technology), and β-actin (12262, Cell Signaling Technology).

Techniques: Derivative Assay, Infection, Injection

Journal: Journal of Neuroinflammation

Article Title: Chi3l3: a potential key orchestrator of eosinophil recruitment in meningitis induced by Angiostrongylus cantonensis

doi: 10.1186/s12974-018-1071-2

Figure Lengend Snippet: Functional protein association networks for the genes correlated with Chi3l3. a A full view of the protein association network for the genes highly correlated with Chi3l3. b The sub-network specifically centered on Chi3l3. c Functional enrichment for the genes in cluster 1 of gene expression patterns. The top 10 biological processes with the observed gene count as well as the false discovery rate are listed. d IL-13 mRNA levels in normal and AC-infected mouse brains. * P < 0.05, AC-infected 21 dpi group vs control group. e A mathematical model for Chi3l3-IL-13 positive feedback and bifurcation analysis. (A, B) Fitting the model to the experimental data. (C, D) Bistability of Chi3l3 and IL-13 with respect to the parameter value ( K 2 ). Data information: In ( d – e ), data are presented as mean ± SD. (Student’s t test)

Article Snippet: The following primary antibodies were used: mouse JMJD3 (ab85392, Abcam), CREB1 (A11989, ABclonal), CEBPB (A0711, ABclonal), KLF4 (A11853, ABclonal), Y641 phosphorylated-STAT6 (ab54461, Abcam), STAT6 (5397, Cell Signaling Technology), Chi3l3 (#60130, STEMCELL Technology), PPARγ (81B8, Cell Signaling Technology), and β-actin (12262, Cell Signaling Technology).

Techniques: Functional Assay, Expressing, Infection

Journal: Journal of Neuroinflammation

Article Title: Chi3l3: a potential key orchestrator of eosinophil recruitment in meningitis induced by Angiostrongylus cantonensis

doi: 10.1186/s12974-018-1071-2

Figure Lengend Snippet: A positive feedback loop between Chi3l3 from macrophages and IL-13 from T lymphocytes in vitro . a The percent of IL-5 + cells in CD3 + CD4 + spleen cells were detected by flow cytometry. b The percent of IL-13 + cells in CD3 + CD4 + cells. * P < 0.05, AC-infected D21 (21 dpi) group vs control group. c , d Spleen cells isolated from normal mice and AC-infected mice were stimulated with sAg (25 μg/mL) or Chi3l3 (10 ng/mL) for 72 h in vitro. A summary of the percentages of IL-13 + CD3 + CD4 + cells following stimulation with sAg and Chi3l3 in normal mice. * P < 0.05, Chi3l3 group vs control group. e Western blot analysis of Chi3l3, JMJD3, CREB1, CEBPB, KLF4, Y641 phospho-STAT6, STAT6, and PPARγ of BMDMs in the presence of sAg, IL-13, sAg+IL-13, and LPS for 24 h. f Cells were cultured for 24 h in medium alone or treated with sAg, IL-13, or sAg+IL-13, and then, the OCR was monitored using the Seahorse Bioscience extracellular flux analyzer in real time. Dotted lines indicate incubation of cells with the indicated compounds. g Basal OCR of BMDMs cultured for 24 h in medium alone or treated with sAg, IL-13, or sAg+IL-13. ** P < 0.01, *** P < 0.001, **** P < 0.0001, sAg group, IL-13 group, sAg+IL-13 group vs control group. # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001, sAg group and IL-13 group vs sAg+IL-13 group. h SRC of BMDMs cultured for 24 h in medium alone or treated with sAg, IL-13, or sAg+IL-13. ** P < 0.01, *** P < 0.001, **** P < 0.0001, sAg group, IL-13 group, sAg+IL-13 group vs control group. # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001, sAg group and IL-13 group vs sAg+IL-13 group. i Maximal OCR of BMDMs cultured for 24 h in medium alone or treated with sAg, IL-13, or sAg+IL-13. ** P < 0.01, *** P < 0.001, **** P < 0.0001, sAg group, IL-13 group, sAg+IL-13 group vs control group. # P < 0.05, ## P < 0.01, ### P < 0.001, #### P < 0.0001, sAg group and IL-13 group vs sAg+IL-13 group. Data information: In ( a – d, f – h ), data are presented as mean ± SD (Student’s t test)

Article Snippet: The following primary antibodies were used: mouse JMJD3 (ab85392, Abcam), CREB1 (A11989, ABclonal), CEBPB (A0711, ABclonal), KLF4 (A11853, ABclonal), Y641 phosphorylated-STAT6 (ab54461, Abcam), STAT6 (5397, Cell Signaling Technology), Chi3l3 (#60130, STEMCELL Technology), PPARγ (81B8, Cell Signaling Technology), and β-actin (12262, Cell Signaling Technology).

Techniques: In Vitro, Flow Cytometry, Infection, Isolation, Western Blot, Cell Culture, Incubation

Journal: Journal of Neuroinflammation

Article Title: Chi3l3: a potential key orchestrator of eosinophil recruitment in meningitis induced by Angiostrongylus cantonensis

doi: 10.1186/s12974-018-1071-2

Figure Lengend Snippet: Top 10 genes ranked according to the maximal changing rate of genes in cluster 1 of gene expression patterns

Article Snippet: The following primary antibodies were used: mouse JMJD3 (ab85392, Abcam), CREB1 (A11989, ABclonal), CEBPB (A0711, ABclonal), KLF4 (A11853, ABclonal), Y641 phosphorylated-STAT6 (ab54461, Abcam), STAT6 (5397, Cell Signaling Technology), Chi3l3 (#60130, STEMCELL Technology), PPARγ (81B8, Cell Signaling Technology), and β-actin (12262, Cell Signaling Technology).

Techniques: Expressing