Journal: PLoS ONE

Article Title: IL-23 Induces Atopic Dermatitis-Like Inflammation Instead of Psoriasis-Like Inflammation in CCR2-Deficient Mice

doi: 10.1371/journal.pone.0058196

Figure Lengend Snippet: On day 12, ( A ) mRNA of indicated cytokines was measured by real-time RT-PCR from ears of WT PBS-injected or WT and CCR2 −/− IL-23-injected mice. Average of 11 mice per genotype in 3 experiments. *p<0.01. ( B ) Intracellular cytokine staining for IL-4 on gated CD3 + CD4 + T cells following stimulation with PMA and ionomycin was performed and day 12 and analyzed by flow cytometry. Number of IL-4 + CD3 + CD4 + T cells in draining lymph node (left panel) or IL-23-injected ear (right panel). ( C ) TSLP was measured by tissue ELISA of ears isolated from PBS-injected or IL-23-injected WT and CCR2 −/− mice. Average of 3 mice per genotype. *p<0.005 CCR2 KO IL-23 vs. all other groups. ( D ) Representative immunofluorescence staining of ear skin isolated on day 12 from IL-23-injected WT C57Bl/6 or CCR2 −/− mice. Sections were stained with Ig isotype control antibody or anti-TSLP (x600, original magnification). Data are reflective of 3 mice per genotype.

Article Snippet: Sections were then washed twice in PBS 0.1% Tween and stained overnight with 1 μg of an affinity purified polyclonal antibody to mouse TSLP (R & D Systems, BAF555) diluted in PBS 0.1% saponin at 4 ° C. After washing with PBS 0.1% Tween on ice, sections were stained with 1 μg of AlexaFluor 488-conjugated donkey anti-goat IgG (Invitrogen) for 1 hour at room temperature.

Techniques: Quantitative RT-PCR, Injection, Staining, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Isolation, Immunofluorescence, Control

Journal: Annals of Dermatology

Article Title: Selective Inhibition of β-Catenin/Co-Activator Cyclic AMP Response Element-Binding Protein-Dependent Signaling Prevents the Emergence of Hapten-Induced Atopic Dermatitis-Like Dermatitis

doi: 10.5021/ad.2019.31.6.631

Figure Lengend Snippet: Effect of ICG-001 on the serum levels of immunoglobulin E (IgE), thymic stromal lymphopoietin (TSLP), thymus and activation-regulated chemokine (TARC), and periostin. ICG-001 treatment of atopic dermatitis (AD) mice suppressed serum TSLP and TARC levels (B, C), and tended to suppress serum IgE and periostin levels, without significance (A, D). Control group (normal) (n=4), Vehicle only mice (oxazolone [Ox]+vehicle) (n=8), ICG-001 treatment (Ox+ICG-001) (n=8). * 0.05> p >0.01, ** p <0.01.

Article Snippet: Levels of TSLP, TARC, periostin and immunoglobulin E (IgE) in serum were examined by ELISA using a mouse TSLP Quantikine ELISA Kit (R&D Systems), a mouse CCL17/thymus and activation-regulated chemokine (TARC) Quantikine ELISA Kit (R&D Systems), a mouse periostin/OSF-2 Quantikine ELISA Kit (R&D Systems), and a mouse IgE quantitation kit (Yamasa Co., Ltd., Chiba, Japan) according to each manufacturer's instructions.

Techniques: Activation Assay, Control

Journal: Mucosal immunology

Article Title: Dendritic cells produce TSLP that limits the differentiation of Th17 cells, fosters Treg development, and protects against colitis.

doi: 10.1038/mi.2011.64

Figure Lengend Snippet: Figure 3 Mouse dendritic cells (DCs) express thymic stromal lymphopoietin (TSLP) after toll-like receptor engagement. ( a , b ) BMDCs were activated with 1 g ml − 1 lipopolysaccharide (LPS), 10 g ml − 1 lipotheicoic acid (LTA), 1 mol l − 1 — C — phosphate — G — (CpG), or 0.1 g ml − 1 flagellin for 3 h. Tslp gene expression was assayed in triplicate by quantitative real-time PCR. Each sample was normalized to Tbp expression. Results represent means ± s.e.m. Data are representative from one of two independent experiments. * P < 0.05, * * P < 0.01, * * * P < 0.001. ( c ) BMDCs were stimulated 3 h with LPS (1 g ml − 1 ) and for 4 h with PMA / ionomicine. Splenic CD11c + DCs were isolated from mice 24 h after intravenous injection with LPS. TSLP was immunoprecipitated from total protein extracts. TSLP expression was detected by western blotting. Protein bands were quantified by densitometry and expressed relative to the untreated sample as arbitrary units. rTSLP, recombinant TSLP; NT, not treated; WT, wild-type.

Article Snippet: In some cultures, different concentrations of recombinant mouse TSLP (R & D Systems) were added (0.2, 1, 5, 10, and 20 ng ml − 1 ).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Expressing, Isolation, Injection, Immunoprecipitation, Western Blot, Recombinant

Journal: Mucosal immunology

Article Title: Dendritic cells produce TSLP that limits the differentiation of Th17 cells, fosters Treg development, and protects against colitis.

doi: 10.1038/mi.2011.64

Figure Lengend Snippet: Figure 4 Dendritic cell-derived thymic stromal lymphopoietin (TSLP) controls Th17 and Treg cell differentiation. 2 × 10 4 BMDCs from C57BL / 6J mice were activated or not with lipopolysaccharide (LPS). After washing, cells were cocultured with 10 5 CD4 + CD25 − T cells from spleens of wild-type (WT) or TSLPR − / − BALB / c mice for 6 days. ( a ) Intracellular cytokine staining was carried out after 6 days of coculture. Left, representative dot plots showing percentages of interferon (IFN)- + and interleukin (IL)-17 + T cells in the CD4 + gate. Right, frequencies of CD4 + IFN- + and CD4 + IL-17 + T cells of three individual experiments are shown. Error bars represent s.e.m. ( b ) After 4 days, cells were stained for CD4 and Foxp3. Percentages of CD4 + Foxp3 + T cells are shown. Error bars represent s.e.m. of triplicate samples. Data are representative from one out of three independent experiments. * P < 0.05, * * P < 0.01, * * * P < 0.001. NS: nonsignificant.

Article Snippet: In some cultures, different concentrations of recombinant mouse TSLP (R & D Systems) were added (0.2, 1, 5, 10, and 20 ng ml − 1 ).

Techniques: Derivative Assay, Cell Differentiation, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: A critical role for thymic stromal lymphopoietin in nickel-induced allergy in mice.

doi: 10.4049/jimmunol.1300276

Figure Lengend Snippet: FIGURE 2. TSLP expression in ear tissues of mice with Ni allergy. (A) Quantitative RT-PCR analysis of Tslp mRNA expression in ear tissues of naive mice and those injected with PBS + CFA (control), Ni (Ni + CFA), Au (Au + CFA), or Pd (Pd + CFA) 24 h after the second treatment. Data are mean 6 SD for 10 mice/group. (B) ELISA of TSLP levels in ear tissue samples (1.5-mm diameter) from mice with Ni allergy and controls 48 h after the second challenge. Data are mean 6 SD for five mice/group. (C) ELISA of TSLP levels in ear tissue samples from mice with Ni allergy 0–48 h after the second treatment. Data are mean 6 SD for five mice/ group. (D) TSLP expression in the allergic lesions was detected using immunofluorescence assays. Results are representative of four mice/group. Alexa Fluor 568–conjugated anti-goat IgG was used as the secondary Ab. Cell nuclei were stained with DAPI. Arrows indicate TSLP expression. The results represent three independent experiments. *p , 0.05.

Article Snippet: To determine TSLP expression, the specimens were incubated with anti-mouse TSLP Ab (R&D Systems) and subsequently with Alexa Fluor 568–conjugated anti-goat IgG (Invitrogen).

Techniques: Expressing, Quantitative RT-PCR, Injection, Control, Enzyme-linked Immunosorbent Assay, Staining

Journal: International Journal of Molecular Sciences

Article Title: Blockade of PAR2 Signaling by Punicalagin as a Therapeutic Strategy for Atopic Dermatitis

doi: 10.3390/ijms26188920

Figure Lengend Snippet: PCG suppresses PAR2-induced inflammatory signaling in HaCaT cells. ( A ) Representative immunoblot showing the effect of PCG on ERK1/2 and p65 phosphorylation. HaCaT cells were pretreated with various concentrations of PCG for 20 min before stimulation with 30 µM PAR2-AP. ( B , C ) Quantification of phosphorylated ERK1/2 and p65 levels. Phosphorylated ERK1/2 bands were normalized to total ERK1/2 and phosphorylated p65 (p-p65) was normalized to total p65. ( D ) Effect of PCG on PAR2-AP-induced IL-8 secretion. HaCaT cells were pretreated with PCG for 0 min and then stimulated with 30 µM PAR2-AP for 6 h. IL-8 levels in cell culture supernatants were measured by ELISA. Data are expressed as mean ± SEM ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The levels of TSLP were measured using a mouse TSLP ELISA kit (catalog #MTLP00; R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Western Blot, Phospho-proteomics, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Blockade of PAR2 Signaling by Punicalagin as a Therapeutic Strategy for Atopic Dermatitis

doi: 10.3390/ijms26188920

Figure Lengend Snippet: PCG normalizes epidermal hyperplasia, inflammatory markers, and neuronal hypersensitivity in DNFB-induced atopic dermatitis. ( A ) Representative H&E-stained skin sections showing epidermal thickness on day 28. Scale bar = 100 µm. ( B ) Quantitative analysis of epidermal thickness measured from H&E-stained sections ( n = 5–6 mice per group). ( C ) Serum TSLP levels measured by ELISA. ND, not detected ( n = 5–6 mice per group). ( D ) Calcium imaging of dorsal root ganglion (DRG) neurons in response to PAR2-AP (5 µM) stimulation. Traces show [Ca 2+ ] i changes over time, with KCl (100 mM) used as a positive control for neuronal viability ( n ≥ 20 per group). Data are presented as mean ± SEM. *** p < 0.001 compared to DNFB group.

Article Snippet: The levels of TSLP were measured using a mouse TSLP ELISA kit (catalog #MTLP00; R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Imaging, Positive Control

Journal: Heliyon

Article Title: Anti-atopic dermatitis effect of fraxinellone via inhibiting IL-31 in vivo and in vitro.

doi: 10.1016/j.heliyon.2024.e35391

Figure Lengend Snippet: Fig. 4. Fraxinellone inhibits the expression of inflammatory cytokines in the lesion skin and serum of DNFB-induced AD-like mice. The levels of IL- 31(A), IL-4 (B), and TSLP (C) in the serum of AD mice were measured by ELISA kit. The levels of IL-31(D), IL-4 (E), and TSLP (F) in the lesion skin in each group (*P < 0.05, **P < 0.01, ***P < 0.001, versus DNFB group; #P < 0.05, ##P < 0.01, ###P < 0.001, comparison between groups; N.S, not significant; n = 4).

Article Snippet: A hematoxylin and eosin (H&E) staining kit (cat: G1120, Solarbio), mouse IL-31 enzyme-linked immunosorbent assay (ELISA) kit (cat: EK1100, Boster), mouse IL-4 ELISA kit (cat: TW10973, Tongwei), mouse Thymic stromal lymphopoietin (TSLP) ELISA kit (cat: E-EL-M0646, Elabscience) were used.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Developmental cell

Article Title: Niche-specific factors dynamically regulate sebaceous gland stem cells in the skin

doi: 10.1016/j.devcel.2019.08.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: All mice were maintained in specific pathogen free housing and were used in accordance with regulations established by the University of Michigan Unit for Laboratory Animal Medicine. table ft1 table-wrap mode="anchored" t5 caption a7 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-androgen receptor Santa Cruz Cat # sc-816 Rabbit anti-cleaved caspase-3 Cell Signaling Cat # 9661 Mouse anti-FASN BD Cat # 610963 Rabbit anti-Gata6 Cell Signaling Cat # D61E4 Chicken anti-GFP Aves Labs Cat # GFP-1010 Rabbit anti-Hes1 Cell Signaling Cat # 11988S Rabbit anti-Ki67 Cell Signaling Cat # 12202S Guinea Pig anti-K5 American Research Products Cat # 03-GP-CK5 Goat anti-K14 Santa Cruz (SC) Cat # sc-17104 Rabbit anti-K14 Covance (COV) Cat # PRB-155P Rabbit anti-K17 Coulombe Lab N/A Goat anti-K79 Santa Cruz Cat # sc-243156 Rabbit anti-K79 (NOTE: see Veniaminova et al., 2013 ) Abcam Cat # Ab7195 Goat anti-Lrig1 R&D Systems Cat # AF3688-SP Rabbit anti-NICD Cell Signaling Cat # 4147P Rabbit anti-Notch1 Cell Signaling Cat # 3608S Rabbit anti-PPARy Santa Cruz Cat # sc-7196 Rabbit anti-Rbpj Cell Signaling Cat # 5313P Goat anti-Tslp R&D Systems Cat # AF555-SP Biological Samples Mouse skin samples, obtained in accordance with guidelines established by the University of Michigan Unit for Laboratory Animal Medicine This manuscript Study protocol # PRO00008196 Chemicals, Peptides, and Recombinant Proteins Tamoxifen Sigma Cat # T5648 TSA Fluorescein Plus kit Perkin Elmer Cat # NEL741E001KT Nile Red Sigma Cat # N3013 RNeasy Mini Kit Qiagen Cat # 74104 Deposited Data RNA-seq data This manuscript GEO: {"type":"entrez-geo","attrs":{"text":"GSE125737","term_id":"125737"}} GSE125737 Experimental Models: Organisms/Strains Mouse: Lrig1 tm1.1(cre/ERT2)Rjc (Lrig1-Cre ERT2 ) Powell et al., 2012 ; The Jackson Laboratory Cat # 018418 Mouse: Tg(KRT14-cre/ERT)20Efu/J (K14-Cre ERT ) Vasioukhin et al., 1999 ; The Jackson Laboratory Cat # 005107 Mouse: Rbpj tm1Hon (Rbp-j flox ) Yamamoto et al., 2003 ; Laboratory of Dr. Tasuku Honjo MGI:3583755 Mouse: Notch1 tm2Rko /GridJ (Notch1 flox ) Yang et al., 2004 ; The Jackson Laboratory Cat # 007181 Mouse: Gt(ROSA)26Sor tm1(Smo/EYFP)Amc /J (SmoM2) Mao et al., 2006 ; The Jackson Laboratory Cat # 005130 Mouse: Krt79 tm2b(KOMP)Wtsi (K79-KO) Mesler et al., 2017 ; Laboratory of Dr.

Techniques: Recombinant, Software