rabbit anti-vti1a  (Synaptic Systems)

Journal: Acta biochimica et biophysica Sinica

Article Title: Tim-1-mediated extracellular matrix promotes the development of hepatocellular carcinoma.

doi: 10.3724/abbs.2024191

Figure Lengend Snippet: Figure 6. Tim-1 enhances cytokine secretion in hepatic stellate cells via NF-κB-related signaling pathways A plasmid construct designed to overexpress Tim-1 was generated and transduced into the JS1 cell line via lentivirus. (A) Fluorescence microscopy examination of Tim-1 protein expression in the JS1 cell line following stable overexpression. (B) Western blot analysis confirmed Tim-1 protein overexpression. (C) Quantification of Tim-1 protein levels in (B) by calculating the relative grayscale values using ImageJ. (D) CCK8 assay was used to evaluate cell proliferation before and after Tim-1 overexpression. These Tim-1-overexpressing cells were analyzed for the levels of intracellular Th1-type TNF-α (E), INF-γ (F), and IL-1β (G) and Th2-type TGF-β (H), MMP2 (I), and IL-4 (J) using corresponding ELISA kits. A comprehensive analysis of the bulk RNA-seq data obtained from JS1 cells displaying high Tim-1 expression was performed (n=3 each). (K,L) Heatmaps showing upregulated and downregulated genes. (M,O) GO enrichment analysis of upregulated and downregulated genes. (N,P) KEGG pathway enrichment analysis of upregulated and downregulated genes. (Q) JS1 cells with high Tim-1 expression were subjected to western blot analysis with antibodies against phosphorylated Akt, phosphorylated NF-κB p65, Akt, and NF-κB p65. (R) Quantification of the relative levels of phosphorylated Akt and phosphorylated NF-κB p65 in (Q) by calculating the relative grayscale values via ImageJ. **P<0.01; ***P<0.001.

Article Snippet: The membrane was incubated overnight at 4°C with specific antibodies, including antibodies against Tim-1 (sc-518008; 1:1000) and COL1A2 (sc-393573; 1:1000) (Santa Cruz Biotechnology, Santa Cruz, USA), phospho-Akt (Ser473) (1:5000; #4060), Akt (1:1000; #9272), phospho-NF-κB p65 (Ser536) (1:1000; #3033), NF-κB p65 (1:1000; #4764), and GAPDH (1:2000; #5174) (Cell Signaling Technology, Danvers, USA).

Techniques: Protein-Protein interactions, Plasmid Preparation, Construct, Generated, Fluorescence, Microscopy, Expressing, Over Expression, Western Blot, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, RNA Sequencing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Hepatic TM6SF2 Is Required for Lipidation of VLDL in a Pre-Golgi Compartment in Mice and Rats

doi: 10.1016/j.jcmgh.2021.12.008

Figure Lengend Snippet: Antibodies Used in the Experiments Described in This Article

Article Snippet: Rabbit anti–hVTI1A pAb , h, m, r , Cell Signaling Technology , 14764 , 1:1000.

Techniques:

Journal: Journal of Inflammation Research

Article Title: LCC-09, a Novel Salicylanilide Derivative, Exerts Anti-Inflammatory Effect in Vascular Endothelial Cells

doi: 10.2147/JIR.S305168

Figure Lengend Snippet: LCC-09 inhibits activation of NFκB and STAT1, and ROS generation in endothelial cells. HUVECs were pre-treated with LCC-09 and control DMSO for 30 mins at the indicated concentrations and then stimulated with TNFα (5 ng/mL). ( A and B ) Cell lysates were subjected to Western blotting. ( C ) ChIP was performed to examine the binding of NFκB to E-selectin promoter 2 h after TNFα stimulation. ( D and E ) Cells were stained with Cellrox Green and imaged ( D ). Fluorescent intensity was measured with a SpectraMax plate reader and compared ( E ) 30 mins after TNFα stimulation. *p<0.05; **p<0.01; ***p<0.001.

Article Snippet: Antibodies against NFκB p65 (#4764, 1:1000), phosphorylated p65 S536 (#3031, 1:1000), STAT1 (#9172, 1:1000), phosphorylated STAT1 Y701 (#9167, 1:1000), STAT3 (#4904, 1:1000), phosphorylated STAT3 Y705 (#9131, 1:1000), JNK (#9258, 1:1000), phosphorylated JNK T183/Y185 (#4668, 1:1000), ERK (#9102, 1:1000), phosphorylated ERK T202/Y204 (#9101, 1:1000), p38 (#8690, 1:1000) and phosphorylated p38 T180/Y182 (#4511, 1:1000), and IκBα (#2682, 1:1000), were purchased from Cell Signaling Technology.

Techniques: Activation Assay, Control, Western Blot, Binding Assay, Staining