Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Multifunctional Co‐Delivery Systems with Downregulation of the Novel Target PIM1 in Macrophages to Ameliorate TF‐Mediated Coagulopathy in Sepsis

doi: 10.1002/smll.202412688

Figure Lengend Snippet: PIM1 inhibitor SMI‐4a protects septic mice against coagulation activation and sepsis‐induced acute lung injury. A) Schematic representation of animal experimental procedures. B) Representative western blot membranes and corresponding densitometric analyses of (C) PIM1 in lung tissue (n = 6/group). D) mRNA levels of PIM1 in murine lung tissues (n = 5/group). E) Platelet count in mice (n = 4/group). F–H) Plasma levels of coagulation‐related factors in mice plasma by ELISA, including (F) TAT, (G) D‐dimer, and (H) fibrinogen (n = 4/group). I) The lung sections were subjected to hematoxylin and eosin (HE) staining, F4/80, fibrin, and PIM1 immunohistochemical analysis (n = 4/group; scale bar:50 µm). J) Representative western blot membranes and corresponding densitometric analyses of (K) TF, (L) PAI‐1, (M) Thrombin in lung tissue (n = 6/group). N) mRNA levels of TF in murine lung tissues (n = 5/group). O–Q) mRNA levels of (O) IL‐1β, (P) IL‐6, and (Q) TNF‐ɑ in murine lung tissues (n = 5/group). R) Survival curves of mice in all groups (n = 10/group). Each bar represents the mean ± SD. Statistical analysis for three or more groups was carried out using one‐way ANOVA (C‐H and K‐Q). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The concentrations of PIM1 (CSB‐ E11825 h, Cusabio, China) in human plasma and TAT (CSB‐ E08433 m, Cusabio, China), Fbg (CSB‐ E08202 m, Cusabio, China), and D2D (CSB‐ E13584 m, Cusabio, China) in mouse plasma were assessed using ELISA kits according to the guidelines outlined in the respective ELISA kits.

Techniques: Coagulation, Activation Assay, Western Blot, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemical staining

Journal: International Journal of Molecular Sciences

Article Title: Role of Autophagy in Von Willebrand Factor Secretion by Endothelial Cells and in the In Vivo Thrombin-Antithrombin Complex Formation Promoted by the HIV-1 Matrix Protein p17

doi: 10.3390/ijms21062022

Figure Lengend Snippet: Increased TAT complex formation is observed in plasma from wild type but not autophagy-deficient mice treated with p17. C57BL/6 ( A – D ) and BECN-1 ( E – H ) mice were i.v. injected or not (NT, not treated mice) with 1 ng/mL ( A , E ), 10 ng/mL ( B , F ), 100 ng/mL ( C , G ) and 250 ng/mL ( D , H ) of p17, respectively. Blood samples were collected at 30 and 60 min following p17 injection and further analyzed for TAT complex formation by ELISA. Results are expressed as TAT concentration in plasma of mice. Statistical analysis was performed by one-way ANOVA with Tukey’s multiple comparison test (** p < 0.01, *** p < 0.001, **** p < 0.0001). In box and whiskers graphs, boxes extend from the 25th to the 75th percentiles, lines indicate the median values, and whiskers indicate the range of values. Each box represents the mean ± SEM of five animals.

Article Snippet: Citrated plasma samples were collected at 30 and 60 min following p17 injection and analyzed for the presence of TAT complexes by ELISA (Assaypro LLC., St. Charles, MO, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Cell Death & Disease

Article Title: YOD1 protects against MRSA sepsis-induced DIC through Lys33-linked deubiquitination of NLRP3

doi: 10.1038/s41419-024-06731-5

Figure Lengend Snippet: A , B Immunoblot analysis of supernatants (SN) or cell lysates (CL) of BMDMs from WT and Yod1 −/− mice, then treated with PBS or MRSA for 4 h. NLRP3, p20 and p17 expression levels were quantitated by measuring band intensities using “ImageJ” software. The values were normalized to β-actin. C , D Representative images of ASC specks in MRSA-infected WT and Yod1 −/− BMDMs. ASC, green; nuclei, blue. Scale bar, 50 μm. The percentage of cells containing an ASC speck was quantified. E – G ELISA of IL-1β ( E ), IL-6 ( F ), and TNF-α ( G ) concentration in supernatants of BMDMs from WT and Yod1 −/− mice after MRSA infection. H , I Immunoblot analysis of liver tissue from WT and Yod1 −/− mice. NLRP3 expression levels were quantitated by measuring band intensities using “ImageJ” software. The values were normalized to β-actin. J ELISA analysis of plasma levels of IL-1β from WT and Yod1 −/− mice after MRSA injection. Data are presented as mean ± SD. ** p < 0.01, *** p < 0.001, **** p < 0.0001, NS means no significance.

Article Snippet: Mouse D-Dimer ELISA Kit (E-EL-M0400c), Mouse thrombin-antithrombin (TAT) ELISA Kit (E-EL-M1138c), Mouse interleukin 1 Beta (IL-1β) ELISA Kit (E-EL-M0037c), Mouse plasminogen activator inhibitor 1 (PAI-1) ELISA Kit (E-EL-M3041) were purchased from Elabscience (Wuhan, China).

Techniques: Western Blot, Expressing, Software, Infection, Enzyme-linked Immunosorbent Assay, Concentration Assay, Clinical Proteomics, Injection