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 reagents were used in this study: Recombinant Murine M-CSF (315-02; Peprotech), Recombinant Mouse Chi3l3 (2446-CH-050, R&D Systems), and Recombinant Mouse IL-13 (413-ML-025/CF, R&D Systems).

Techniques: Expressing, Infection, Control, Gene Expression

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 reagents were used in this study: Recombinant Murine M-CSF (315-02; Peprotech), Recombinant Mouse Chi3l3 (2446-CH-050, R&D Systems), and Recombinant Mouse IL-13 (413-ML-025/CF, R&D Systems).

Techniques: Derivative Assay, Infection, Control, Saline, 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 reagents were used in this study: Recombinant Murine M-CSF (315-02; Peprotech), Recombinant Mouse Chi3l3 (2446-CH-050, R&D Systems), and Recombinant Mouse IL-13 (413-ML-025/CF, R&D Systems).

Techniques: Functional Assay, Gene Expression, Infection, Control

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 reagents were used in this study: Recombinant Murine M-CSF (315-02; Peprotech), Recombinant Mouse Chi3l3 (2446-CH-050, R&D Systems), and Recombinant Mouse IL-13 (413-ML-025/CF, R&D Systems).

Techniques: In Vitro, Flow Cytometry, Infection, Control, 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 reagents were used in this study: Recombinant Murine M-CSF (315-02; Peprotech), Recombinant Mouse Chi3l3 (2446-CH-050, R&D Systems), and Recombinant Mouse IL-13 (413-ML-025/CF, R&D Systems).

Techniques: Gene Expression, Expressing

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: HSP70-binding motifs function as protein quality control degrons

doi: 10.1007/s00018-022-04679-3

Figure Lengend Snippet: The APPY peptide functions as a degron in human cells. a Upon transfection of the HEK293T landing pad cells, BxbI catalyzes site-specific recombination between BxbI attachment sites in the vector and the landing pad, leading to single-copy expression of GFP-APPY from the Tet-on promoter. As the vector also contains an internal ribosomal entry site (IRES) followed by mCherry, the GFP-APPY levels can be normalized to the mCherry levels. 2A signifies self-cleaving peptides. b GFP:mCherry ratio distributions of cells expressing the indicated GFP-APPY variants, obtained by flow-cytometry of ~ 10,000 cells. c GFP:mCherry ratio distributions of cells expressing the GFP-APPY (RLLL) peptide or the DAAA variant of APPY after 5 h treatment with 15 μM bortezomib (BZ) or DMSO (solvent control), obtained by flow-cytometry of ~ 10,000 cells. d GFP:mCherry ratio distributions of cells expressing the APPY (RLLL) peptide or the DAAA variant of APPY after 22 h treatment with 5 μM YM01 or no treatment, obtained by flow-cytometry of ~ 10,000 cells. e Relative expression of the HSPA1A and HSPA1B genes derived from qPCR. The results were normalized to the expression of non-recombinant cells. n = 3, error bars show the standard deviation, asterisks indicate significant differences (p < 0.05) based on a two-tailed unpaired Student’s t test

Article Snippet: HEK293T drug treatments and flow cytometry profiling For the YM01 treatment cells were treated with 5 μM of YM01 (StressMarq Biosciences) for 22 h before flow-cytometry profiling.

Techniques: Transfection, Plasmid Preparation, Expressing, Flow Cytometry, Variant Assay, Solvent, Control, Derivative Assay, Recombinant, Standard Deviation, Two Tailed Test

Journal: Bioscience Reports

Article Title: Atractylon treatment prevents sleep-disordered breathing-induced cognitive dysfunction by suppression of chronic intermittent hypoxia-induced M1 microglial activation

doi: 10.1042/BSR20192800

Figure Lengend Snippet: ( A–D ) qRt-PCR detection of the expression of M1 macrophage markers (IFN-γ and iNOS) and M2 macrophage markers (Arg1 and Ym1). Data are presented as the mean ± SD. ** P <0.01, *** P <0.001 versus the control group. ### P <0.001 versus CIH treatment group.

Article Snippet: The following primary Abs were used: monoclonal Ab against GAPDH (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, U.S.A.), SIRT3 (1:600; Cell Signaling Technology), nuclear factor (NF)-κB (1:600; Santa Cruz Biotechnology), interferon-γ (IFN-γ; 1:600; Santa Cruz Biotechnology), arginase-1 (Arg1; 1:600; Santa Cruz Biotechnology), inducible nitric oxide synthase (iNOS; 1:600; Santa Cruz Biotechnology), chitinase 3-like 3 (Ym1; 1:600; Santa Cruz Biotechnology), and Iba-1 (1:600; Santa Cruz Biotechnology).

Techniques: Quantitative RT-PCR, Expressing, Control

Journal: Bioscience Reports

Article Title: Atractylon treatment prevents sleep-disordered breathing-induced cognitive dysfunction by suppression of chronic intermittent hypoxia-induced M1 microglial activation

doi: 10.1042/BSR20192800

Figure Lengend Snippet: ( A–E ) Western blot detection of the effect of SIRT3 on the expression levels of the pro-inflammatory M1 molecules IFN-γ and iNOS, and the M2 molecules Arg1 and Ym1 after exposure to CIH. Data are presented as the mean ± SD. *** P <0.001 versus the control group. ### P <0.001 versus the CIH treatment group.

Article Snippet: The following primary Abs were used: monoclonal Ab against GAPDH (1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, U.S.A.), SIRT3 (1:600; Cell Signaling Technology), nuclear factor (NF)-κB (1:600; Santa Cruz Biotechnology), interferon-γ (IFN-γ; 1:600; Santa Cruz Biotechnology), arginase-1 (Arg1; 1:600; Santa Cruz Biotechnology), inducible nitric oxide synthase (iNOS; 1:600; Santa Cruz Biotechnology), chitinase 3-like 3 (Ym1; 1:600; Santa Cruz Biotechnology), and Iba-1 (1:600; Santa Cruz Biotechnology).

Techniques: Western Blot, Expressing, Control