recombinant mouse protein s pros1  (R&D Systems)

Journal: Comprehensive Physiology

Article Title: Porcine Fetal Hypothyroidism Induces Temporal and Tissue‐Specific Alterations in the Insulin‐Like Growth Factor System

doi: 10.1002/cph4.70028

Figure Lengend Snippet: IGFBP levels in fetal serum. (A) Example Western Ligand Blot showing human recombinant protein standards in the left lane, with the right lane showing the six bands detected in fetal porcine serum. Relative density of (B) a 26 kDa IGFBP, (C) a 28 kDa IGFBP, (D) IGFBP2, (E) the 40 kDa IGFBP3, and (F) IGFBP4 in fetal porcine serum derived from fetuses ( n = 11–12/group) at various gestational timepoints following 21 days of maternal CON or MMI treatment. Relative density was calculated relative to a consistent reference sample run on each gel, with unique letter superscripts denoting statistically significant ( p < 0.05) changes in density in the CON group over time, and statistical differences between the CON and MMI group within each timepoint denoted as * p < 0.05, ** p < 0.01, and *** p < 0.001. Data are presented as the estimated marginal means with 95% confidence intervals, as extracted from a linear model including gestational day, treatment group, and the interaction.

Article Snippet: In addition to samples of interest, each blot contained a lane of pooled recombinant human protein standards representing IGFBPs ‐1, ‐2, ‐3, and ‐4 (R&D Systems), which were used as reference points for identification of bands of interest, as well as a lane of pooled fetal serum which was used to calculate the relative density of each band of interest for analysis.

Techniques: Western Blot, Recombinant, Derivative Assay

Journal: Cancers

Article Title: MICA+ Tumor Cell Upregulated Macrophage-Secreted MMP9 via PROS1-AXL Axis to Induce Tumor Immune Escape in Advanced Hepatocellular Carcinoma (HCC).

doi: 10.3390/cancers16020269

Figure Lengend Snippet: Figure 4. MICA+ tumor cells communicated with MMP9+ macrophages via the PROS1-AXL and CCL15-CCR1 pathways. (a,b) The interaction net count and interaction weight plot of HCC cells. The thicker line indicates the higher number of interactions and the stronger weights/strength of interactions between the two cell types. (c) Chord diagram showing all the significant interactions (ligand-receptor pairs) between MICA+/MICA−tumor cells and MMP9+/MMP9−macrophage. (d) The violin plot showing PROS1-AXL/CCL15-CCR1 (ligand-receptor pairs) and some macrophage marker genes expression in different cell clusters. (e) PROS1-AXL/CCL15-CCR1 ligand-receptor pairs and some major IFN-γ signaling pathway genes inside tumor cells and macrophage. Diamonds represent genes that are upregulated in macrophages. Octagons represent genes that are upregulated in tumor cells. The triangles represent transcription factors.

Article Snippet: In certain experiments, the cells were treated with phorbol-12-myristate-13-acetate (PMA) (MedChemExpress, Monmouth Junction, NJ, USA, catalog no. HY-18739/CS-6053), human recombinant Pros1 protein (R&D Systems, Minneapolis, MN, USA, catalog no. 9489-PS) or cabozantinib (MedChemExpress, Monmouth Junction, NJ, USA, catalog no. HY-13016).

Techniques: Marker, Expressing

Journal: Cancers

Article Title: MICA+ Tumor Cell Upregulated Macrophage-Secreted MMP9 via PROS1-AXL Axis to Induce Tumor Immune Escape in Advanced Hepatocellular Carcinoma (HCC).

doi: 10.3390/cancers16020269

Figure Lengend Snippet: Figure 6. IRF1 upregulated PROS1 in HCC cells and also enhanced PROS1 expression in MICA+ HCC cells. (a) The typical IHC images of MICA, MMP9, PROS1, and AXL in non-tumor and tumor live (×400 magnification). Scale bars are 200 µm. (b,c) PROS1 expression levels of IRF1 and MICA positive tumor cells in scRNA-seq data. (d) PROS1 mRNA expression levels of Huh-7 cells infected by lentivirus IRF1 cDNA or empty vector (NC) (50 MOI) for 48 h. (e) PROS1 protein level of Huh-7 cells infected by lentivirus IRF1 cDNA or empty vector (NC) (50 MOI) for 48 h. The quantitative analyses are shown on the right. (f) PROS1 mRNA expression level of Huh-7 cells infected by lentivirus MICA cDNA or empty vector (NC) (50 MOI) for 48 h. (g) PROS1 protein level of Huh-7 cells infected by lentivirus MICA cDNA or empty vector (NC) (50 MOI) for 48 h. The quantitative analyses are shown on the right. (h,i) The typical IF images show PROS1 protein level (red staining) in Huh-7 cells which were respectively transfected with lentiviral IRF1 and MICA (×200 magnification). Scale bars are 150 µm. Quantification of PROS1 via relative fluorescent intensity is shown. ** p < 0.01, *** p < 0.001, via two-tailed unpaired t-test. The original uncropped western blot images are shown in Supplementary Figure S5.

Article Snippet: In certain experiments, the cells were treated with phorbol-12-myristate-13-acetate (PMA) (MedChemExpress, Monmouth Junction, NJ, USA, catalog no. HY-18739/CS-6053), human recombinant Pros1 protein (R&D Systems, Minneapolis, MN, USA, catalog no. 9489-PS) or cabozantinib (MedChemExpress, Monmouth Junction, NJ, USA, catalog no. HY-13016).

Techniques: Expressing, Infection, Plasmid Preparation, Staining, Transfection, Two Tailed Test, Western Blot

Journal: Cancers

Article Title: MICA+ Tumor Cell Upregulated Macrophage-Secreted MMP9 via PROS1-AXL Axis to Induce Tumor Immune Escape in Advanced Hepatocellular Carcinoma (HCC).

doi: 10.3390/cancers16020269

Figure Lengend Snippet: Figure 7. MICA+ HCC cell stimulated MMP9 secretion via PROS1-AXL axis and promoted proteolytic shedding of MICA into soluble MICA (sMICA). (a) MMP9, AXL, CD163 and CD206 mRNA expression was detected via qPCR in Huh-7 cells stimulated by recombinant human PROS1 protein 2 µg/mL for 72 h. (b) The whole lysate for PROS1 and AXL protein determined via western blot is shown in macrophage stimulated by recombinant human PROS1 protein for 72 h. The quantitative analyses are shown on the right. (c,d) The level of MMP9 and soluble MICA in the conditioned medium was determined via an ELISA procedure. (e) MMP9 mRNA expression was detected via qPCR in macrophage co-cultured with MICA+ and NC Huh7 cells. The whole lysate for MMP9 protein determined via western blot is shown in macrophage cultured with MICA+ and NC Huh7 cells. The quantitative analyses are shown on the right. (f) MMP9 mRNA expression was detected via qPCR in macrophage co-cultured with IRF1+ and NC Huh7 cells. The whole lysate for MMP9 protein determined via western blot is shown in macrophage cultured by IRF1+ and NC Huh7 cells. The quantitative analyses are shown on the right. (g) MMP9 protein expression was measured via western blot in macrophage stimulated by preconditioned cabozantinib (5 µM) and recombinant human PROS1 protein for 72 h. The quantitative analyses are shown on the right. (h) MMP9 protein expression was measured via western blot in macrophage co-cultured with IRF1+ and NC Huh7 cells treated by cabozantinib (5 µM). The quantitative analyses are shown on the right. * p < 0.05, ** p < 0.01, *** p < 0.001, via two-tailed unpaired t-test. The original western blots are shown in Supplementary Figure S5.

Article Snippet: In certain experiments, the cells were treated with phorbol-12-myristate-13-acetate (PMA) (MedChemExpress, Monmouth Junction, NJ, USA, catalog no. HY-18739/CS-6053), human recombinant Pros1 protein (R&D Systems, Minneapolis, MN, USA, catalog no. 9489-PS) or cabozantinib (MedChemExpress, Monmouth Junction, NJ, USA, catalog no. HY-13016).

Techniques: Expressing, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Cell Culture, Two Tailed Test

Journal: Cancers

Article Title: MICA+ Tumor Cell Upregulated Macrophage-Secreted MMP9 via PROS1-AXL Axis to Induce Tumor Immune Escape in Advanced Hepatocellular Carcinoma (HCC).

doi: 10.3390/cancers16020269

Figure Lengend Snippet: Figure 8. System illustration of MICA+ tumor cell upregulated macrophage-secreted MMP9 via PROS1-AXL axis to induce tumor immune escape in advanced HCC. (a) IRF1 transcriptionally upregulated MICA expression to activate NKG2D-mediated anti-tumor immune responses by CD8+ T cells. (b) DNA damage induced MICA and PROS1 expression via IRF1. PROS1 activated MMP9 secreted from macrophages via AXL receptor to hydrolyze MICA. The release of soluble MICA (sMICA) promoted immune evasion by the tumor.

Article Snippet: In certain experiments, the cells were treated with phorbol-12-myristate-13-acetate (PMA) (MedChemExpress, Monmouth Junction, NJ, USA, catalog no. HY-18739/CS-6053), human recombinant Pros1 protein (R&D Systems, Minneapolis, MN, USA, catalog no. 9489-PS) or cabozantinib (MedChemExpress, Monmouth Junction, NJ, USA, catalog no. HY-13016).

Techniques: Expressing