Journal: BMC pharmacology & toxicology

Article Title: Blocking group 2 innate lymphoid cell activation and macrophage M2 polarization: potential therapeutic mechanisms in ovalbumin-induced allergic asthma by calycosin.

doi: 10.1186/s40360-024-00751-9

Figure Lengend Snippet: Fig. 2 Calycosin inhibited group 2 innate lymphoid cell (ILC2) activation in OVA-induced asthmatic mice. (A) The levels of interleukin (IL)-4, (B) IL-5 and (C) IL-13 in the BALF and lung tissues were measured using ELISA and RT-qPCR. (D) ELISA was used to determin the contents of IL-33 in the BALF. (E) Suppression tumorigenicity 2 (ST2) levels in the lung tissues was examined by western blot. The blots were cut prior to hybridization with antibodies during blotting. (F) Immunohistochemistry (IHC) staining was used to detect the expression ST2 and IL-25R. (G) The co-localization of ST2 and CD45 was detected by immunofluorescence (IF). Scale bar = 50 μm. n = 6 for each group. * P< 0.05, **P < 0.01, ***P < 0.001, compared with the sham group; #P < 0.05, ##P < 0.01, ###P < 0.001, compared with the OVA group. “Ca” represents calycosin

Article Snippet: The sections were incubated with ST2 antibody (1:50; 11920- 1-AP, ProteinTech, Wuhan, China) at 4°C overnight and subjected to HRP-labeled goat anti-rabbit IgG (1:500; # 31460, ThermoFisher Scientific, USA) at 37°C for 1 h. Additionally, IL-25R expression was detected in the same way.

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Hybridization, Immunohistochemistry, Expressing, Immunofluorescence

Journal: BMC pharmacology & toxicology

Article Title: Blocking group 2 innate lymphoid cell activation and macrophage M2 polarization: potential therapeutic mechanisms in ovalbumin-induced allergic asthma by calycosin.

doi: 10.1186/s40360-024-00751-9

Figure Lengend Snippet: Fig. 4 ILC2 activation induced by IL-33 was restrained by calycosin in ILC2s. (A) The ILC2 cell viability was examined by CCK-8. (B) Western blot showed the expression of ST2 in ILC2s. The blots were cut prior to hybridization with antibodies during blotting. (C) The secretion of IL-4, (D) IL-5 and (E) IL-13 in ILC2s were detected by ELISA assay. n = 3 for each group. **P < 0.01,***P < 0.001, compared with the control group; #P < 0.05, ##P < 0.01, ###P < 0.001, compared with the IL-33 group. “Ca” represents calycosin

Article Snippet: The sections were incubated with ST2 antibody (1:50; 11920- 1-AP, ProteinTech, Wuhan, China) at 4°C overnight and subjected to HRP-labeled goat anti-rabbit IgG (1:500; # 31460, ThermoFisher Scientific, USA) at 37°C for 1 h. Additionally, IL-25R expression was detected in the same way.

Techniques: Activation Assay, CCK-8 Assay, Western Blot, Expressing, Hybridization, Enzyme-linked Immunosorbent Assay, Control

Journal: Inflammation and Regeneration

Article Title: Novel artificial nerve transplantation of human iPSC-derived neurite bundles enhanced nerve regeneration after peripheral nerve injury

doi: 10.1186/s41232-024-00319-4

Figure Lengend Snippet: Early-stage tissue evaluation after neurite bundle-derived artificial nerve transplantation. a , b Immunohistochemistry images with anti-NFH and anti-CD31 antibodies. Dotted arrows indicate the proximal side of the intact nerve areas, and solid arrows indicate the NFH- and CD31-positive regeneration tip in the reconstructed tissue. Scale bars = 1 mm. c , d Quantitative comparison of axonal extension distance was indicated by NFH-positive areas and that of vascular elongation distance was demonstrated by CD31-positive areas at 1 and 2 weeks after transplantation. Significant differences between groups elongation distances were observed in 2-week samples only. e , f Iba1 immunohistochemistry image of the proximal part of the regenerated tissue at 2 weeks after transplantation (e). Scale bars = 1 mm. Quantitative analysis of Iba1-positive areas demonstrated that a larger number of macrophages aggregated in the Auto, motor TP and sensory TP (f). ( n = 3). * p < 0.05, ** p < 0.01, N.S. = not significant. Data are represented as the mean ± SEM

Article Snippet: The primary antibodies used in this study were human polyclonal anti-pan-ELAVL (ELAV-like protein 2/3/4) antibody (1:2000, kindly provided from Prof. Robert Darnell, Rockefeller University), rabbit polyclonal anti-TrkA antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), chicken polyclonal anti-TrkB antibody (1:500, kindly provided from Prof. Louis F. Reichardt, UCSF), goat polyclonal anti-TrkC antibody (AF373, 1:200, R&D systems), rabbit polyclonal anti-parvalbumin antibody (PV-28, 1:1000, Swant), rabbit polyclonal anti-CGRP antibody (BML-CA1134, 1:500, Enzo Life Sciences), Goat polyclonal anti-Choline acetyltransferase antibody (AB144P, 1:200, Chemicon), Mouse monoclonal anti-Islet1 and Islet2 antibody (39.4D5, 1:200, DSHB), rabbit polyclonal anti-Neurofilament heavy polypeptide antibody (ab8135, 1:500, Abcam), goat polyclonal anti-mouse and anti-rat CD31 (AF3628, 1:100, R&D), rabbit polyclonal anti-Iba1 antibody (GtX100042, 1:500, GeneTex), goat anti-GFP antibody (600–101-215, 1:1000, Rockland).

Techniques: Derivative Assay, Transplantation Assay, Immunohistochemistry, Comparison

Journal: Journal of Nanobiotechnology

Article Title: Momordica charantia L.-derived exosome-like nanovesicles stabilize p62 expression to ameliorate doxorubicin cardiotoxicity

doi: 10.1186/s12951-024-02705-z

Figure Lengend Snippet: MC-ELNs prevent p62 from DOX-induced ubiquitination degradation in cardiomyocytes. H9c2 cells were pre-treated with 10 µg/mL MC-ELNs for 6 h, and then cultured with or without 1 µM doxorubicin (DOX) for another 24 h. Western blot images ( A ) and quantitative analysis ( B ) on the p62 protein levels. ( C ) The p62 mRNA levels in H9c2 cells treated with 1 µM DOX in the absence or presence of 10 µg/mL MC-ELNs. ( D ) H9c2 cells were treated with PBS or 1 µM DOX in the presence or absence of 2 µM Lactacystin (LACTA) for 24 h. ( E ) H9c2 cells were incubated in the absence or presence of MC-ELNs (10 µg/mL) in combination with the protein synthesis inhibitor cycloheximide (CHX, 10 µg/mL) for 2–4 h. Western blot analysis of the expressions of p62. ( F ) Immunoprecipitation assay (IP) and western blot analysis of cell lysate derived from PBS- or DOX-treated H9c2 cells in the absence or presence of 10 µg/mL of MC-ELNs (DOX, DOX + MC-ELNs). IP was performed with an antibody against p62. The blot was probed with antibodies against p62, Ub, K48-Ub, and K63-Ub. β-actin was employed as a loading control. Western blot analysis on the identified E3 ligase enzyme bound with p62 including Parkin ( G ), TRIM21 ( H ), and TRAF6 ( I ) in H9c2 cells treated with 1 µM DOX in the absence or presence of 10 µg/mL MC-ELNs. All data are presented as mean ± SD ( n = 3 experiments per group for A - E , G - I ; n = 1 experiment per group for F ). Comparisons among more than two groups were performed by ordinary one-way analysis of variance (ANOVA) followed by the Tukey’s multiple comparisons test

Article Snippet: The primary antibodies included Cleaved caspase3 (9664S, CST), Caspase3 (9665S, CST), Cleaved caspase7 (8438S, CST), Caspase7 (12827T, CST), Cleaved PARP (9545, CST), PARP (9532S, CST), γ-H2A.X (ab2893, Abcam), Cyclin D1 (2978S, CST), Cyclin E1 (20808S, CST), Cyclin B1 (4138S, CST), Nrf2 (SAB4501984, Sigma), HO-1 (10701-1AP, proteintech), SQSTM1/p62 (ab109012, Abcam), Keap1 (8047S, CST), Ubiquitin (ab140601, Abcam), K48-Ub (ab140601, Abcam), K63-Ub (ab179434, Abcam), TRIM21 (ab207728, Abcam), TRAF6 (HY-P80919, MCE), Parkin (ab77924, Abcam), PCNA (ab29, Abcam), LC3B (NB100-2220, NOVUS), NQO1 (67240-1-lg, proteintech), Catalase (21260-1-AP, proteintech), SOD2 (24127-1-AP, proteintech), GPX4 (ab125066, Abcam), β-actin (HRP-600008, proteintech).

Techniques: Ubiquitin Proteomics, Cell Culture, Western Blot, Incubation, Immunoprecipitation, Derivative Assay, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Role and mechanism of quercetin via the TLR4/MyD88/IRAK4 signaling pathway in the treatment of allergic rhinitis

doi: 10.3892/etm.2026.13215

Figure Lengend Snippet: Effect of QUE on mRNA of TLR4, MyD88, IRAK4 and NF-κB. Results of (A) TLR4, (B) MyD88, (C) IRAK4 and (D) NF-κB mRNA relative expression levels in NC, OVA, DEX and QUE groups. Data are presented as the mean ± SEM of three repeats. *** P<0.001, **** P<0.0001 vs. NC; ## P<0.01, ### P<0.001, #### P<0.0001 vs. OVA. NC, normal control; OVA, ovalbumin; DEX, dexamethasone; QUE, quercetin; TLR4, Toll-like receptor 4; MyD88, myeloid differentiation primary response 88; IRAK4, interleukin-1 receptor-associated kinase 4.

Article Snippet: AF8187 and AF7524, respectively; Beyotime Biotechnology), NF-κB p65 (cat. no. 8242; Cell Signaling Technology, Inc.), IRAK4 (cat. no. ab119942, Abcam), BCA kit (Thermo Fisher Scientific, Inc.), SYBR Green qPCR Master Mix and RT Master Mix for quantitative PCR (qPCR; MedChemExpress), ELISA kit OVA-IgE (cat. no. MOFI01017; Assay Genie), mouse IL-17 ELISA Kit (cat. no. CSB-E04608m; Cusabio Technology, LLC), mouse IL-1β ELISA Kit, mouse IL-10 ELISA Kit, mouse IL-13 ELISA Kit, mouse IL-4 ELISA Kit (cat. nos.

Techniques: Expressing, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Role and mechanism of quercetin via the TLR4/MyD88/IRAK4 signaling pathway in the treatment of allergic rhinitis

doi: 10.3892/etm.2026.13215

Figure Lengend Snippet: Effects of QUE on TLR4, MyD88, IRAK4 and NF-κB protein levels. (A) A significant increase was observed in the relative expression of TLR4, MyD88, IRAK4 and NF-κB proteins in the OVA group compared with the NC group. The relative expression of these proteins was lower in the DEX and QUE groups than in the OVA group. Semi-quantification of (B) TLR4, (C) MyD88, (D) IRAK4 and (E) NF-κB protein expression levels. Data are presented as the mean ± SEM of three repeats. * P<0.05, ** P<0.01 vs. NC; # P<0.05, ### P<0.001 vs. OVA. NC, normal control; OVA, ovalbumin; DEX, dexamethasone; QUE, quercetin; TLR4, Toll-like receptor 4; MyD88, myeloid differentiation primary response 88; IRAK4, interleukin-1 receptor-associated kinase 4.

Article Snippet: AF8187 and AF7524, respectively; Beyotime Biotechnology), NF-κB p65 (cat. no. 8242; Cell Signaling Technology, Inc.), IRAK4 (cat. no. ab119942, Abcam), BCA kit (Thermo Fisher Scientific, Inc.), SYBR Green qPCR Master Mix and RT Master Mix for quantitative PCR (qPCR; MedChemExpress), ELISA kit OVA-IgE (cat. no. MOFI01017; Assay Genie), mouse IL-17 ELISA Kit (cat. no. CSB-E04608m; Cusabio Technology, LLC), mouse IL-1β ELISA Kit, mouse IL-10 ELISA Kit, mouse IL-13 ELISA Kit, mouse IL-4 ELISA Kit (cat. nos.

Techniques: Expressing, Control

Journal: Structure (London, England : 1993)

Article Title: Functional Relevance of IL-1 Receptor Inter-domain Flexibility for Cytokine Binding and Signaling

doi: 10.1016/j.str.2019.05.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit Anti-IL-1RAcP Antibody , Rockland , Cat# 600–401-BV3; RRID:AB_2613201.

Techniques: Recombinant, Plasmid Preparation, Software

Journal: Bone

Article Title: Type 2 innate lymphoid cells inhibit the differentiation of osteoclasts and protect from ovariectomy-induced bone loss.

doi: 10.1016/j.bone.2020.115335

Figure Lengend Snippet: Fig. 1. ILC2 directly suppress osteoclastogenesis in vitro. (a) Bone marrow-derived monocytes (BMM) were co-cultured with innate lymphoid cells type 2 (ILC2; ILC2:BMM ratio = 1:10). Osteoclasts were stained by tartrate acid resistant phosphatase (TRAP) after stimulation with 50 ng/mL RANKL for 5 days. Scale bar = 200 μm, Representative data are from one out of three in- dependent experiments. (b) TRAP-positive cells were counted (Nuclei ≥3). (c) Pit formation assay. BMM were cultured with ILC2 (ILC2:BMM ratio = 1:20 or 1:10). Representative data are from one out of three independent experiments. Scale bar = 200 μm (d) Measurement of resorption pit area. (e) Immunohistochemistry using mouse femur bone sample. Osteoclasts (white arrows with asterisk) were stained by ELF97 substrate and ILC2 (white arrows) were labeled as ST2+IL17RB+lineage (CD3e, CD11b, CD11c, B220, Ly6G) negative cells (n = 6). Scale bar = 50 μm. Data are expressed as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: ILC2 were stained using antibodies against IL17RB (Santa Cruz, sc11754), ST2 (ProSci, 3363), CD3ε (eBioscience, 17-0031), CD11b (R&D systems, 1124), CD11c (Abcam, ab33483), B220 (eBioscience, 14-0452), Ly6G (R&D systems, MAB1037), and DAPI.

Techniques: In Vitro, Derivative Assay, Cell Culture, Staining, Tube Formation Assay, Immunohistochemistry, Labeling

Journal: Bone

Article Title: Type 2 innate lymphoid cells inhibit the differentiation of osteoclasts and protect from ovariectomy-induced bone loss.

doi: 10.1016/j.bone.2020.115335

Figure Lengend Snippet: Fig. 3. Adoptive transfer of ILC2 inhibits osteoclasts in vivo. (a) Light sheet microscopy image of tibia. 1 × 106 sort-purified ILC2 (red cells with white arrows) were antibody labeled and intravenously injected into recipient mice. Asterisks indicate bone tissue. Scale bar indicates 1000 μm. (b) Frequencies of innate lymphoid cells type 2 (ILC2) in the spleen after adoptive transfer. ILC2 were identified as CD5−B220−CD45R−NKp46−CD11b−CD11c−ICOS+ KLRG1+ ST2+ cells. (c) Frequencies of T-bet+ ILC1 and NCR+/−Rorgt+ ILC3 in the spleen of mice with and without adoptive transfer of ILC2s. (d) Histomorphometry of tibial bones showing the number of osteoclasts per bone area (N.Oc/T.Ar) and osteoclast number per bone perimeter (N.Oc/B.Pm) with and without adoptive transfer of ILC2s. (e) μCT analysis of tibial bones showing bone volume per tissue volume (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N) and trabecular separation (Tb.Sp) with and without adoptive transfer of ILC2s. (f) Frequencies of T-bet+Th1 cells, Gata3+Th2 cells, Rorgt+Th17 cells and FoxP3+T cells in the spleen of mice with and without adoptive transfer of ILC2s. Data are from two independent experiments. Data are expressed as mean ± SEM. *p < 0.05; **p < 0.01. n = 4–5/group.

Article Snippet: ILC2 were stained using antibodies against IL17RB (Santa Cruz, sc11754), ST2 (ProSci, 3363), CD3ε (eBioscience, 17-0031), CD11b (R&D systems, 1124), CD11c (Abcam, ab33483), B220 (eBioscience, 14-0452), Ly6G (R&D systems, MAB1037), and DAPI.

Techniques: Adoptive Transfer Assay, In Vivo, Microscopy, Purification, Labeling, Injection