Journal: bioRxiv

Article Title: MNS induces antiviral protection and suppresses inflammation

doi: 10.64898/2026.02.11.705318

Figure Lengend Snippet: (A-C) IL1B mRNA (qPCR) and protein levels (ELISA) in RAW 264.7 cells (A), BMDMs (B), and THP1 cells (C) following treatment with DMSO or MNS and infection with indicated viruses. (D-G) Il1b expression (qPCR) and protein levels (ELISA) in liver tissues from C57BL/6J mice under viral challenge and MNS treatment. (H) MNS markedly downregulated the expression of Il1b , Il6 , Tnf α, Ptgs2 , Nos2 mRNA in RAW264.7 cells. (I) MNS significantly reduced the expression of COX2, iNOS, p-IκBα, p-P65 at the protein level. (J) MNS treatment led to a significant reduction in mortality among LPS-challenged mice (n=13). (K) The expression level of Il1b in heart, spleen, lung, kidney, and liver was quantified by RT-qPCR. Values are presented as mean ± SEM. Significance was indicated as follows: N.S., p > 0.05; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: Mouse cells were assayed with a mouse IL1B ELISA kit (Boster, EK2285), and human primary cells were assayed with a human IL1B ELISA kit (Boster, EK2286), following the manufacturers’ instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Infection, Expressing, Quantitative RT-PCR

Journal: iScience

Article Title: 17β-estradiol and estrogen receptor alpha protect mouse ovarian follicle development by repressing atresia

doi: 10.1016/j.isci.2025.111846

Figure Lengend Snippet: Detection of apoptotic cells and FSH levels in the ovaries and sera of mice administered eCG or OVA (A–F) Apoptotic cells were detected using the TUNEL assay with sections prepared from the ovaries from mice 12 h (A and B), 24 h (C and D), and 48 h (E and F) after eCG or OVA injection. Enlarged images in the box in (A–C) and (E) are shown in (A′–C′) and (E′), respectively. The area surrounded by the line and dotted line in (A′–C′) and (E′) represent follicle and oocyte (Oo), respectively. Arrowheads indicate the follicles with apoptotic GCs. Scale bars: 200 μm (A, C, E); 100 μm (B, D, F); 50 μm (A′, B′, C′, E′). (G) The apoptotic index was expressed as the percentage of the TUNEL-positive cells area per whole area of the follicle. ∗∗ p < 0.01 (ANOVA and Tukey-Kramer test, n = 7). (H) Plasma FSH levels were measured using ELISA in mice 12 and 48 h after eCG or OVA injection (t test, n = 4).

Article Snippet: Activin A, FSH, E 2 , and testosterone levels were measured using a Mouse Activin A ELISA Kit PicoKine® (BOSTER Biological Technology, Pleasanton, CA), a Mouse FSH ELISA Kit (MyBioSource, Inc., San Diego, CA), an Estradiol ELISA Kit, and a Testosterone ELISA Kit, respectively, according to the manufacturer’s instructions.

Techniques: TUNEL Assay, Injection, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: 17β-estradiol and estrogen receptor alpha protect mouse ovarian follicle development by repressing atresia

doi: 10.1016/j.isci.2025.111846

Figure Lengend Snippet: INHA and INHBA expression in ovaries and activin A and inhibin A levels in sera of mice administered eCG or OVA (A) INHA and INHBA expression were detected by western blotting using extracts prepared from the ovaries from mice 12 h and 48 h after eCG or OVA injection. (B and C) ACTB was used as a loading control. Plasma inhibin A and/or inhibin B (B) and activin A (C) levels were measured with ELISA using sera collected from mice administered eCG or OVA. ∗∗ p < 0.01, ∗ p < 0.05 (t test, n = 4).

Article Snippet: Activin A, FSH, E 2 , and testosterone levels were measured using a Mouse Activin A ELISA Kit PicoKine® (BOSTER Biological Technology, Pleasanton, CA), a Mouse FSH ELISA Kit (MyBioSource, Inc., San Diego, CA), an Estradiol ELISA Kit, and a Testosterone ELISA Kit, respectively, according to the manufacturer’s instructions.

Techniques: Expressing, Western Blot, Injection, Control, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: 17β-estradiol and estrogen receptor alpha protect mouse ovarian follicle development by repressing atresia

doi: 10.1016/j.isci.2025.111846

Figure Lengend Snippet: Expression of steroidogenic enzyme genes and E 2 levels in ovaries and sera of mice administered eCG or OVA (A and B) RT-qPCR analysis of steroidogenic enzyme gene expression using cDNA prepared from the ovaries of mice collected 12 h (A) and 48 h (B) after eCG or OVA injection. ∗ p < 0.05 (t test, n = 5). (C) CYP19A1 expression was detected by western blotting of extracts prepared from mouse ovaries collected 12 and 48 h after eCG or OVA injection. ACTB was used as a loading control (two upper panels). The signal intensity of the bands in the upper panels was quantified using densitometric analysis, and the ratio of CYP19A1 to ACTB expression was calculated as the relative expression (lower panels). ∗ p < 0.05 and ∗∗ p < 0.01 (t test, n = 4). (D) E 2 and testosterone levels were measured in the sera collected from mice 12, 24, and 48 h after eCG or OVA injection using ELISA. ∗ p < 0.05, ∗∗ p < 0.01 (t test, n = 6). (E) E 2 and testosterone levels were measured in the ovaries of mice collected 12 and 48 h after eCG or OVA injection using ELISA. ∗ p < 0.05 (t test, n = 7).

Article Snippet: Activin A, FSH, E 2 , and testosterone levels were measured using a Mouse Activin A ELISA Kit PicoKine® (BOSTER Biological Technology, Pleasanton, CA), a Mouse FSH ELISA Kit (MyBioSource, Inc., San Diego, CA), an Estradiol ELISA Kit, and a Testosterone ELISA Kit, respectively, according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Gene Expression, Injection, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: 17β-estradiol and estrogen receptor alpha protect mouse ovarian follicle development by repressing atresia

doi: 10.1016/j.isci.2025.111846

Figure Lengend Snippet: Effect of activin A on Cyp19a1 and Esr expression and E 2 production in cultured mouse ovaries (A, C, and D) The expression of Cyp19a1 (A), Esr1 (C), and Esr2 (D) were examined using RT-qPCR using cDNA prepared from ovaries cultured with or without activin A. ∗ p < 0.05 (t test, n = 3). (B) E 2 levels in the culture media were measured using ELISA. ∗∗ p < 0.01 (t test, n = 3).

Article Snippet: Activin A, FSH, E 2 , and testosterone levels were measured using a Mouse Activin A ELISA Kit PicoKine® (BOSTER Biological Technology, Pleasanton, CA), a Mouse FSH ELISA Kit (MyBioSource, Inc., San Diego, CA), an Estradiol ELISA Kit, and a Testosterone ELISA Kit, respectively, according to the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: 17β-estradiol and estrogen receptor alpha protect mouse ovarian follicle development by repressing atresia

doi: 10.1016/j.isci.2025.111846

Figure Lengend Snippet:

Article Snippet: Activin A, FSH, E 2 , and testosterone levels were measured using a Mouse Activin A ELISA Kit PicoKine® (BOSTER Biological Technology, Pleasanton, CA), a Mouse FSH ELISA Kit (MyBioSource, Inc., San Diego, CA), an Estradiol ELISA Kit, and a Testosterone ELISA Kit, respectively, according to the manufacturer’s instructions.

Techniques: Virus, Recombinant, TUNEL Assay, In Situ, Enzyme-linked Immunosorbent Assay, Software

Journal: Cell host & microbe

Article Title: Leukotriene B4-Mediated Neutrophil Recruitment Causes Pulmonary Capillaritis during Lethal Fungal Sepsis.

doi: 10.1016/j.chom.2017.11.009

Figure Lengend Snippet: Figure 2. Rapid PMN Intravascular Chemotaxis to Sequestered C. albicans Is Complement Dependent (A) Lung C5a levels were determined using ELISA in control and C3/ animals, following i.v. C. albicans. Two-way ANOVA, Sidak’s multiple comparison. (B) Using intravital microscopy, PMN chemotactic behaviors in C3/ and anti-C5aR mAb-treated mice were observed. Images shown are 10 min after C. albicans injection. Red arrows highlight yeast not recognized by PMN. Scale bar represents 30 mm. (C) Quantification of PMN chemotaxing to pathogens in C3/ and C5a receptor-blocked mice. One-way ANOVA, Dunnett’s multiple comparison. (D) The percentage of C. albicans phagocytosed during the initial 10 min of intravenous administration in control, C3/, and anti-C5aR mAb-treated mice using pulmonary intravital microscopy. t tests were used to compare C57BL/6 and anti-C5aR mAb treated (*) or C3/ (#) mouse time points. (E and F) C. albicans CFU was quantified in organs at (E) 1 hr and (F) 24 hr after injection with 1 3 106 C. albicans in mice treated with anti-C5a receptor antibodies or isotype. (G) Clinical sepsis scores of isotype and anti-C5aR mAb antibody treated mice 24 hr after injection with 1 3 106 C. albicans. A score of 21 is the clinical endpoint for euthanization. For (A)–(F), at least three individual experiments were performed; for (G) three individual experiments were performed. Error bars represent mean ± SEM. *p < 0.05, **p < 0.01.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-Ly6G (clone 1A8) BioLegend RRID: AB_2563207 (BioLegend Cat. No. 127636) Anti-CD31 (clone MEC13.3) BioLegend RRID: AB_2161030 (BioLegend Cat. No. 102515) Anti-CD45 (clone 30-F11) BioLegend RRID: AB_493532 (BioLegend Cat. No. 103121) Anti-CD49b (clone HMa2) BD Bioscience Catalog No. 558295 Anti-C5aR (CD88, clone 20/70) BioLegend RRID: AB_2067286 (BioLegend Cat. No. 135804) Purified anti-mouse Ly6G (clone 1A8) BioXCell BP0075-1 Anti-TER119 Biolegend RRID: AB_528961 (BioLegend Cat. No. 116218) Anti-human CD15 (clone W6D3) BioLegend RRID: AB_756018 (BioLegend Cat. No. 323012) Anti-human CD45 (clone 2D1) R&D Systems Catalog # FAB1430G Anti-human C5aR (CD88, clone S5/1) BioLegend RRID: AB_2259318 (BioLegend Cat. No. 344302) Bacterial and Virus Strains C. albicans (strain ATCC58716) ATCC LUMC-101 Chemicals, Peptides, and Recombinant Proteins Zymosan A (S. cerevisiae) BioParticles ThermoFisher Z23373 Escherichia coli (K-12 strain) BioParticles ThermoFisher E13231 SYTO 9 Green Fluorescent Nucleic Acid Stain ThermoFisher S34854 Compstatin Tocris Cat. No. 2585 LY293111 Cayman Chemical Company CAS N 161172-51-6 Recombinant mouse C5a R&D Systems 2150-C5-025 Critical Commercial Assays LTB4 Parameter Assay Kit R&D Systems KGE006B EasySep Mouse Biotin Positive Selection Kit STEMCELL Technologies Catalog # 18556 Mouse Complement C5a PicoKine ELISA Kit Boster Biological Technology EK0987 Polydimethylsiloxane (PDMS) microfluidic devices SynvivoBio Cat # 104001 Experimental Models: Cell Lines human lung microvascular endothelial cells HLMEC Lonza CC-2527 Experimental Models: Organisms/Strains C57BL/6J The Jackson Laboratory Stock No: 000664 LTB4R knockout (B6.129S4-Ltb4r1tm1Adl/J) The Jackson Laboratory Stock No: 008102 C3 / ; (B6;129S4-C3tm1Crr/J) Gift from Paul Kubes N/A CD11b / ; (B6.129S4-Itgamtm1Myd/J) Gift from Paul Kubes N/A CD11a / ; (B6.129S7-Itgaltm1Bll/J) Gift from Paul Kubes N/A LysM-GFP Gift from Paul Kubes N/A Ly6G-cre (C57BL/6-Ly6g(tm2621(CretdTomato)Arte)) Gift from Matthias Gunzer N/A Software and Algorithms Volocity PerkinElmer N/A Leica LAS Leica N/A Prism 7 software GraphPad V7.02 FlowJo FlowJo V10.1 e1 Cell Host & Microbe 23, 1–13.e1–e4, January 10, 2018

Techniques: Chemotaxis Assay, Enzyme-linked Immunosorbent Assay, Control, Comparison, Intravital Microscopy, Injection

Journal: Frontiers in Pharmacology

Article Title: CD226 Is Required to Maintain Megakaryocytes/Platelets Homeostasis in the Treatment of Knee Osteoarthritis With Platelet-Rich Plasma in Mice

doi: 10.3389/fphar.2021.732453

Figure Lengend Snippet: Deletion of platelet CD226 disturbed the homeostasis of MKs/platelets. (A) Identification of CD226 fl/fl and CD226 fl/fl PF4-Cre mice through PCR of DNAs extracted from the tail. (B) Flow cytometry images showing CD226 expression in CD226 fl/fl and CD226 fl/fl PF4-Cre mice. (C) HE staining of BM and spleen sections. The MKs are circled; scale bar = 50 µm. (D, E) MK number and average MK area in BM and spleen sections. (F) Routine blood test results of platelets (PLT) number, plateletcrit (PCT), mean platelet volume (MPV), platelet distribution width (PDW), platelet-large cell ratio (P-LCR), and platelet-large cell count (P-LCC). Statistical significance between groups was analyzed using independent-sample t -tests with two-tailed p value. * p < 0.05, ** p < 0.01, ns = no statistical significance.

Article Snippet: The serum concentrations of PDGF‐AB and platelet factor 4 (PF4) were analyzed using their corresponding ELISA kits (#EK0486 for PDGF-AB, #EK0727 for PF4; Boster, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Flow Cytometry, Expressing, Staining, Cell Counting, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: CD226 Is Required to Maintain Megakaryocytes/Platelets Homeostasis in the Treatment of Knee Osteoarthritis With Platelet-Rich Plasma in Mice

doi: 10.3389/fphar.2021.732453

Figure Lengend Snippet: Absence of CD226 in platelets hindered platelet maturation and reduces α-granule secretion. (A) Platelet number was evaluated by flow cytometry. (B) Percentage of TO positive and negative platelets was measured to show the maturation of platelets. Two-ANOVA was adopted for statistical analysis over time. (C) Morphological changes of platelets. Red arrow: α‐granule; blue arrow: dense-granule. (D) Area of platelet, area of total α‐granule per platelet, and average area of total α‐granule per platelet were calculated. (E) Concentrations of PDGF-AB and PF4 in serum. Statistical significance was analyzed by independent-sample t -tests with two-tailed p value. * p < 0.05, *** p < 0.001, ns = no statistical significance.

Article Snippet: The serum concentrations of PDGF‐AB and platelet factor 4 (PF4) were analyzed using their corresponding ELISA kits (#EK0486 for PDGF-AB, #EK0727 for PF4; Boster, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Flow Cytometry, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: CD226 Is Required to Maintain Megakaryocytes/Platelets Homeostasis in the Treatment of Knee Osteoarthritis With Platelet-Rich Plasma in Mice

doi: 10.3389/fphar.2021.732453

Figure Lengend Snippet: Platelet-specific CD226 absence led to abnormal ribosomal function and structure. (A) A GO analysis of the DEPs. (B) Annotation of the gene ID. (C) Volcano plot of differentially expressing proteins from the CD226 fl/fl PF4-Cre platelet versus littermates ( n = 3 per group).

Article Snippet: The serum concentrations of PDGF‐AB and platelet factor 4 (PF4) were analyzed using their corresponding ELISA kits (#EK0486 for PDGF-AB, #EK0727 for PF4; Boster, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Expressing

Journal: Frontiers in Pharmacology

Article Title: CD226 Is Required to Maintain Megakaryocytes/Platelets Homeostasis in the Treatment of Knee Osteoarthritis With Platelet-Rich Plasma in Mice

doi: 10.3389/fphar.2021.732453

Figure Lengend Snippet: CD226 deficiency in platelets decreased the ribosome and autophagy related protein expression in MK/platelets and human Dami cells. (A) S6 ribosomal protein-expressing ribosome in isolated MKs from BM. Scale bar = 50 µm. (B) Lysates of freshly isolated mice platelets were subjected to western blot with anti-S6 ribosomal, PDGF-A, and PDGF-B antibodies. (C) Double immunofluorescence staining for vWF and Beclin1 in the BM slides from CD226 fl/fl PF4-Cre and CD226 fl/fl mice. Scale bar = 50 µm. (D) Lysates of freshly isolated mice platelets were subjected to western blot with anti-LC3, Beclin1, and CD226 antibodies. (E) Dami cells infected with lentivirus shCD226 were subjected to western blot with anti-LC3, Beclin 1, and CD226 antibodies.

Article Snippet: The serum concentrations of PDGF‐AB and platelet factor 4 (PF4) were analyzed using their corresponding ELISA kits (#EK0486 for PDGF-AB, #EK0727 for PF4; Boster, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Expressing, Isolation, Western Blot, Double Immunofluorescence Staining, Infection

Journal: Frontiers in Pharmacology

Article Title: CD226 Is Required to Maintain Megakaryocytes/Platelets Homeostasis in the Treatment of Knee Osteoarthritis With Platelet-Rich Plasma in Mice

doi: 10.3389/fphar.2021.732453

Figure Lengend Snippet: Histological scoring of PRP-treated DMM mice. (A) OARIS-modified Mankin score of articular cartilage. (B) Osteophyte size scores and (C) Osteophyte maturity scores. The score-related data were analyzed by Kruskal–Wallis test accompanied with Dunn’s multiple comparison test. * p < 0.05, ** p < 0.01, ns = no statistical significance. . Flow cytometry analysis for the expression of CD226 in splenocyte from CD226 fl/fl and CD226 fl/fl PF4-Cre mice. (A) Gating strategy for flow cytometric analysis of splenocyte subsets. (B) Representative image of CD226 expression levels in B cells, T cells, NK cells, CD11b + myeloid cells and CD11c + DC. . Other parameter detected after platelet CD226 deficiency in mice. (A) Routine blood test of the hemoglobin (HGB), red blood cell (RBC) number, and white blood cell (WBC) number. (B) Area of total dense-granule per platelet and average area of dense-granule per platelet in two groups. Statistical significance between groups was analyzed using independent-sample t -tests with two-tailed p value. ns = no statistical significance.

Article Snippet: The serum concentrations of PDGF‐AB and platelet factor 4 (PF4) were analyzed using their corresponding ELISA kits (#EK0486 for PDGF-AB, #EK0727 for PF4; Boster, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Modification, Comparison, Flow Cytometry, Expressing, Two Tailed Test