Journal: Molecular Medicine

Article Title: Exploring the mechanism of endothelial Pim-1 upregulation of tissue factor to initiate the hypercoagulable state in sepsis

doi: 10.1186/s10020-026-01433-4

Figure Lengend Snippet: Inhibition of Pim-1 activity increased the survival rate of CLP-induced septic mice (WT) and alleviated the coagulation response. a Immunohistochemical staining (the staining agent was horseradish peroxidase, HRP) and Pim-1 + positive cells statistical plot of WT mice. b Pim-1 protein bands and statistical results in lung/liver of CLP WT mice ( n ≥ 3/group). c Dual immunofluorescence staining images of Pim-1 and CD31 in lung tissues were obtained using Cy3-conjugated goat anti-rabbit IgG (red) and Alexa Fluor 488-conjugated goat anti-mouse IgG (green). d Pim-1 protein bands and statistical results after LPS stimulation of HUVECs. e Survival of experimental mice. WT mice aged 6–8 weeks were given CLP, and six hours later SMI-4a (15 mg/kg or 30 mg/kg) was injected via the tail vein ( n = 10/group; Kaplan-Meier survival analysis). f Plasma TM, TAT, FIB and D-dimer contents of WT mice ( n ≥ 3/group), CLP + SMI-4a (L) : 15 mg/kg, CLP + SMI-4a (H) : 30 mg/kg. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns represent no statistical significance

Article Snippet: We contracted Cyagen Biosciences Inc. to construct a mouse model of specific endothelial cell Pim-1 knockout (KO).

Techniques: Inhibition, Activity Assay, Coagulation, Immunohistochemical staining, Staining, Immunofluorescence, Injection, Clinical Proteomics

Journal: Molecular Medicine

Article Title: Exploring the mechanism of endothelial Pim-1 upregulation of tissue factor to initiate the hypercoagulable state in sepsis

doi: 10.1186/s10020-026-01433-4

Figure Lengend Snippet: Pim-1 relies on the transcription factor Sp1 to regulate TF in HUVECs stimulated with LPS. a Protein bands and statistical plots of p-Sp1 and TF after Pim-1 kinase inhibition in HUVECs (SMI-4a 80 µM). b Protein bands and statistical plots of p-Sp1 with increasing degree of Pim-1 kinase inhibition in HUVECs (SMI-4a 30, 60, 90, 120 µM). c Protein bands and statistical plots of p-Sp1 and TF after LPS stimulation in HUVEs transfected with Pim-1 overexpression plasmid. d Protein bands of TF after HUVECs treated with Pim-1 overexpression plasmid and 200 nM Mithramycin A (Sp1 inhibitor). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns represent no statistical significance

Article Snippet: We contracted Cyagen Biosciences Inc. to construct a mouse model of specific endothelial cell Pim-1 knockout (KO).

Techniques: Inhibition, Transfection, Over Expression, Plasmid Preparation

Journal: Molecular Medicine

Article Title: Exploring the mechanism of endothelial Pim-1 upregulation of tissue factor to initiate the hypercoagulable state in sepsis

doi: 10.1186/s10020-026-01433-4

Figure Lengend Snippet: Pim-1 activates the mTOR/Sp1/TF axis to initiate SIC in LPS-induced HUVECs. a Protein bands and statistical plots of p-mTOR after Pim-1 overexpression in HUVECs. b Protein bands and statistical plots of p-Sp1 and TF after mTOR inhibition in HUVECs (RAPA 5 µM). c - d Protein bands and statistical plots of p-Sp1 and TF after transfection of HUVECs with mTOR overexpression plasmid. e Protein bands of p-Sp1 and TF after HUVEC treated with mTOR overexpression plasmid and 80 µM SMI-4a. **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns represent no statistical significance

Article Snippet: We contracted Cyagen Biosciences Inc. to construct a mouse model of specific endothelial cell Pim-1 knockout (KO).

Techniques: Over Expression, Inhibition, Transfection, Plasmid Preparation

Journal: Molecular Medicine

Article Title: Exploring the mechanism of endothelial Pim-1 upregulation of tissue factor to initiate the hypercoagulable state in sepsis

doi: 10.1186/s10020-026-01433-4

Figure Lengend Snippet: Pim-1 endothelial cell-specific knockout affects coagulation function after CLP-induced sepsis in mice. a and b Images of lung/liver tissues from Pim-1 fl/fl and Pim-1 fl/fl Cre + group subjected to H&E staining and fibrin immunohistochemical staining ( n ≥ 3/group; The staining agents were Hematoxylin-Eosin and HRP; magnification 200×, Scale bar = 10 μm, black arrow pointing to microvascular thrombus). c The blood perfusion volume of the right kidney and mesentery in four groups of mice ( n ≥ 3/group). d Plasma TM, TAT, FIB and D-D dimer levels were measured in Pim-1 fl/fl and Pim-1 fl/fl Cre + group ( n ≥ 3/group). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns represent no statistical significance

Article Snippet: We contracted Cyagen Biosciences Inc. to construct a mouse model of specific endothelial cell Pim-1 knockout (KO).

Techniques: Knock-Out, Coagulation, Staining, Immunohistochemical staining, Clinical Proteomics

Journal: Molecular Medicine

Article Title: Exploring the mechanism of endothelial Pim-1 upregulation of tissue factor to initiate the hypercoagulable state in sepsis

doi: 10.1186/s10020-026-01433-4

Figure Lengend Snippet: Pim-1 endothelial cell-specific knockout affects coagulation function and signaling pathways after CLP-induced sepsis in mice. a and b Protein was extracted from lung tissue of Pim-1 fl/fl and Pim-1 fl/fl Cre + group for immunofluorescence to observe TF and Thrombin protein expression ( n ≥ 3/group), Scale bar = 50 μm. c Protein was extracted from lung tissue of Pim-1 fl/fl and Pim-1 fl/fl Cre + group for western blot to observe TF and Thrombin protein expression ( n ≥ 3/group). d and e Bands and statistical plots of mouse p-mTOR and p-Sp1 proteins in Pim-1 fl/fl and Pim-1 fl/fl Cre + group ( n ≥ 3/group). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns represent no statistical significance

Article Snippet: We contracted Cyagen Biosciences Inc. to construct a mouse model of specific endothelial cell Pim-1 knockout (KO).

Techniques: Knock-Out, Coagulation, Protein-Protein interactions, Immunofluorescence, Expressing, Western Blot

Journal: Molecular Medicine

Article Title: Exploring the mechanism of endothelial Pim-1 upregulation of tissue factor to initiate the hypercoagulable state in sepsis

doi: 10.1186/s10020-026-01433-4

Figure Lengend Snippet: TLR4 affects the expression of Pim-1 and TF in CLP/LPS-induced sepsis. a Immunofluorescence images and statistical graphs of Pim-1 protein in lung tissues of WT group and TLR4 mut group ( n ≥ 3/group), Scale bar = 50 μm. b Immunofluorescence images and statistical graphs of TF protein in lung tissues of WT group and TLR4 mut group ( n ≥ 3/group), Scale bar = 50 μm. c Bands and statistics of Pim-1 protein in lung tissues of WT and TLR4 mut groups ( n ≥ 3/group). d Bands and statistics of TF protein in lung tissues of WT and TLR4 mut groups ( n ≥ 3/group). e Bands and statistical analysis of Pim-1 and TF proteins after inhibition of TLR4 in HUVECs (5 nM TAK-242). f Protein expression and statistical graph of Pim-1 and TF after HUVECs were stimulated with different concentrations of LPS (2.5, 5, 10 μg/ml; LPS is a TLR4 activator). **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, ns represent no statistical significance

Article Snippet: We contracted Cyagen Biosciences Inc. to construct a mouse model of specific endothelial cell Pim-1 knockout (KO).

Techniques: Expressing, Immunofluorescence, Inhibition