crth2  (Proteintech)

Journal: Science Advances

Article Title: CHI3L1/YKL-40 signaling inhibits neurogenesis in models of Alzheimer’s disease

doi: 10.1126/sciadv.adv1492

Figure Lengend Snippet: ( A ) Immunoblots of 17 candidate mediators after 10-min CHI3L1 exposure (0 to 500 ng/ml). Phospho/total signals were densitometrically normalized to the no-CHI3L1 control (1.0); n = 3 to 4. ( B ) Dose-response blots for p-S6K1 and p-S6 (10 min). Values are phospho/total, expressed relative to untreated cells (1.0); n = 4. ( C ) Dose-response blots for p-IKKβ with the same quantification as in (B); n = 3. ( D ) BMS345541 (10 μM, 30 min) was added 10 min before CHI3L1 (500 ng/ml). p-S6 was quantified as in (B); n = 4. ( E ) iNPCs transfected with CRTH2 siRNA (siCRTH2) or control siRNA (siNC) were exposed to CHI3L1 (500 ng/ml, 48 hours). Proliferation = EdU + /DAPI + cells (% of total DAPI + ); n = 4. ( F ) After the same transfection, cells were differentiated for 3 days under CHI3L1 incubation. Neuronal differentiation output = DCX + /DAPI + (%), expressed as fold change over siNC+Ctrl (1.0); n = 4. Statistics: one-way ANOVA with Tukey’s post hoc for [(A) to (C)] and (E); unpaired t test for (D). * P < 0.05; ** P < 0.005, *** P < 0.001; **** P < 0.0001. n.s., not significant.

Article Snippet: To determine the requirement of CRTH2 in the CHI3L1 signaling pathway, we performed RNAi-mediated knockdown of CRTH2 in iNPCs using a validated siRNA targeting human CRTH2 (siCRTH2; Santa Cruz Biotechnology, #sc-39838) or a control siRNA (siCtrl; Santa Cruz Biotechnology, #sc-37007).

Techniques: Western Blot, Control, Transfection, Incubation

Journal: Science Advances

Article Title: CHI3L1/YKL-40 signaling inhibits neurogenesis in models of Alzheimer’s disease

doi: 10.1126/sciadv.adv1492

Figure Lengend Snippet: ( A ) Design and timeline. Lentiviruses expressing shCRTH2-eGFP or shCtrl-eGFP were injected bilaterally into the DG of 6-month-old wild-type (WT) and 5XFAD mice (shCtrl in one side and shCRTH2 in the other). BrdU was given immediately, EdU 24 hours before euthanasia 4 weeks later. Groups: WT + shCtrl, 5XFAD + shCtrl, and 5XFAD + shCRTH2 ( n = 5). ( B ) Thioflavin-S/Aβ 42 staining showed hippocampal plaques in 5XFAD but not WT; CRTH2 knockdown did not change plaque size or density. ( C ) GFAP and CHI3L1 Immunolabeling indicated comparable astrocytosis and CHI3L1 induction in 5XFAD hemispheres with or without shCRTH2. ( D ) SGZ proliferation. EdU+ GFP+ cells were classified as radial NSCs (GFAP+) or amplifying progenitors (GFAP−). Proliferation was reduced in 5XFAD + shCtrl versus WT but significantly restored by shCRTH2. ( E ) Neuronal output. GFP+ BrdU+ cells coexpressing DCX (immature) or NeuN (mature) were decreased in 5XFAD + shCtrl and rescued by CRTH2 knockdown. Scale bars, 100 μm. ** P < 0.005, *** P < 0.001, and **** P < 0.0001 by one-way ANOVA with Tukey’s post hoc (comparisons versus WT + shCtrl and 5XFAD + shCtrl).

Article Snippet: To determine the requirement of CRTH2 in the CHI3L1 signaling pathway, we performed RNAi-mediated knockdown of CRTH2 in iNPCs using a validated siRNA targeting human CRTH2 (siCRTH2; Santa Cruz Biotechnology, #sc-39838) or a control siRNA (siCtrl; Santa Cruz Biotechnology, #sc-37007).

Techniques: Expressing, Injection, Staining, Knockdown, Immunolabeling

Journal: The Journal of Clinical Investigation

Article Title: ICOS regulates IL-10 production in group 2 innate lymphoid cells via cholesterol and cortisol biosynthesis

doi: 10.1172/JCI193134

Figure Lengend Snippet: ( A – H ) hILC2s (CD45 + , lineage – , CRTH2 + , and CD127 + ) were freshly isolated from PBMCs of healthy donors and cultured with or without anti-ICOS ligand (anti-ICOSL). Right panel shows hILC2 purity after being sorted. ( B – F ) Levels of IL-10 ( B ), IL-4 ( C ), IL-5 ( D ), IL-6 ( E ), and IL-13 ( F ) in the culture supernatants following treatment with or without anti-ICOSL. n = 6. ( G and H ) The expression levels of intranuclear Ki67 ( G ) and GATA-3 ( H ) expression is presented as MFI. n = 6. ( I and J ) Representative histogram of MAF ( I ) and NFIL3 ( J ) protein expression levels . Corresponding quantitation is presented as MFI. n = 4. Data are presented as mean ± SEM. Two-tailed Student’s t test was employed for statistical analysis; * P < 0.05, ** P < 0.01, and *** P < 0.001. Schematic images were created in Adobe Illustrator. FMO, fluorescence minus one.

Article Snippet: After red blood cell lysis (BioLegend), CRTH2 + cells were isolated using the CRTH2 MicroBead Kit (Miltenyi Biotec) as per the manufacturer’s protocol.

Techniques: Isolation, Cell Culture, Expressing, Quantitation Assay, Two Tailed Test, Fluorescence

Journal: Advanced Science

Article Title: Coactosin‐Like Protein Reduces Prostaglandin D 2 Production in Alveolar Macrophages and Alleviates Allergic Airway Inflammation

doi: 10.1002/advs.202501673

Figure Lengend Snippet: Effect of reconstitution of clodronate‐treated mice with littermate control or Cotl1 −/‐ alveolar macrophages on HDM‐induced pulmonary immune cell accumulation. A) WT mice received clodronate liposomes to deplete endogenous AMs. After 72 h, the mice were intranasally injected with WT or Cotl1 −/‐ AMs and treated with HDM. On day 23, mice were sacrificed, and lung tissue single‐cell suspensions were prepared and analyzed by flow cytometry. B) Frequencies of IRF5 + M1 macrophages, CD206 + M2 macrophages, and IL‐10 + M2‐like macrophages among the CD45 + CD11c + CD11b − F4/80 + population ( n = 6). C) Frequencies of IL‐4 + Th2 cells among the CD4 + population, CD49b + basophils among the IgE + population, Siglec‐F + eosinophils among the CD45 + CD11c + population, Sca‐1 + KLRG1 + ILC2 cells among the CD45 + CD25 + Lin − population, and CD117 + mast cells among the IgE + population ( n = 6). D) Frequencies of CRTH2 + cells among the macrophage population ( n = 6). E) Frequencies of CRTH2 + cells among the basophil population ( n = 6). F) Frequencies of CRTH2 + cells among the eosinophil population ( n = 6). G) Frequencies of CRTH2 + cells among the ILC2 population ( n = 6). H) Frequencies of CRTH2 + cells among the mast cell population ( n = 6). I) Frequencies of CRTH2 + cells among the Th2 cell population ( n = 6). Data shown in (B), (C), and (E)‐(I) were presented as median ± interquartile range. p values were assessed using Tukey's multiple comparisons test following one‐way ANOVA for (B), (C), and (E–I). p < 0.05 was considered statistically significant.

Article Snippet: Antibodies for CLP (10781‐1‐AP), HPGDS (22522‐1‐AP), mPGES‐2 (10881‐1‐AP), CRTH2 (25264‐1‐AP), and β‐actin (60008‐1‐lg) were purchased from Proteintech (Hubei, China).

Techniques: Control, Liposomes, Injection, Flow Cytometry

Journal: Advanced Science

Article Title: Coactosin‐Like Protein Reduces Prostaglandin D 2 Production in Alveolar Macrophages and Alleviates Allergic Airway Inflammation

doi: 10.1002/advs.202501673

Figure Lengend Snippet: AMs from Cotl1 −/− mice confer exacerbated airway inflammation via CRTH2. A) WT mice received clodronate liposomes to deplete endogenous AMs. After 72 h, the mice were intranasally injected with Cotl1 −/‐ AMs. Mice were challenged with HDM and treated with OC000459 . On day 23, mice were euthanized, and lung tissue was analyzed. B) The levels of Th2 cytokines (IL‐4, IL‐5, and IL‐13) in lung tissue extracts were determined by ELISA ( n = 6). C,D) Representative lung samples stained with (C) H&E and (D) PAS ( n = 6). Scale bar: 50 µm. E) Frequencies of IL‐4 + Th2 cells among the CD4 + population, CD49b + basophils among the IgE + population, Siglec‐F + eosinophils among the CD45 + CD11c + population, Sca‐1 + KLRG1 + ILC2 cells among the CD45 + CD25 + Lin − population, and CD117 + mast cells among the IgE + population ( n = 6). Data shown in (B–D) were presented as mean ± SD, and data shown in (E) were presented as median ± interquartile range. p values were assessed using Tukey's multiple comparisons test following two‐way ANOVA for (B), and Tukey's multiple comparisons test following one‐way ANOVA for (C–E). p < 0.05 was considered statistically significant.

Article Snippet: Antibodies for CLP (10781‐1‐AP), HPGDS (22522‐1‐AP), mPGES‐2 (10881‐1‐AP), CRTH2 (25264‐1‐AP), and β‐actin (60008‐1‐lg) were purchased from Proteintech (Hubei, China).

Techniques: Liposomes, Injection, Enzyme-linked Immunosorbent Assay, Staining