Journal: Oncotarget

Article Title: Chimeric antigen receptor-modified T Cells inhibit the growth and metastases of established tissue factor-positive tumors in NOG mice

doi: 10.18632/oncotarget.14367

Figure Lengend Snippet: ( A ) Structure diagram of TF-CAR. mlF VII, the light chain of mouse coagulation factor VII; TMD, transmembrane domain. ( B ) Western bloting analysis of TF-CAR expression in T cells. CAR-T, TF-CAR T cells; CON-T, vector-transduced T cells; T, untreated T cells. ( C ) Flow cytometry analysis of TF-CAR expression in transduced T cells. ( D ) Immunofluorescence staining analysis of TF-CAR expression in transduced T cells. The images were taken under × 400 magnification. ( E ) CAR positive ratio analysis of lentivirus infected T cells by Flow cytometry. All sample were prepared after 5 days of virus infection.

Article Snippet: The following antigens and antibodies were used for this analysis: human TF, mouse anti-human TF primary antibody (R&D Systems, Minnesota, USA) and FITC-conjugated anti-mouse IgG1 secondary antibody (eBioscience, San Diego, CA); mouse TF, PE-conjugated goat anti-mouse TF antibody (R&D Systems, Minnesota, USA); TF-CAR, polyclonal goat anti-mouse coagulation factor VII primary antibody (R&D Systems, Minnesota, USA) and PE-conjugated rabbit anti-goat IgG secondary antibody (Santa Cruz, CA, USA).

Techniques: Coagulation, Western Blot, Expressing, Plasmid Preparation, Flow Cytometry, Immunofluorescence, Staining, Infection, Virus

Journal: Molecular Metabolism

Article Title: Two-step regulation by matrix Gla protein in brown adipose cell differentiation

doi: 10.1016/j.molmet.2024.101870

Figure Lengend Snippet: Global MGP deficiency disrupts brown adipogenic maturation . ( A ) Expression of the mesenchymal stem cell markers Cd44 , Cdh2 , Vim and Gata2 (left panel) and the adipose progenitor cell markers Cd34 , Cd142 , Icam1 and Pdgfra (right panel), in wild-type (WT) and Mgp -knockout (KO) iBAT, as determined by qPCR (n = 7 mice per group). ( B ) FACS analysis of adipose progenitor cells from the brown adipose stromal vascular fractions isolated from WT and KO mice (n = 5 mice per group), using antibodies to ICAM1, CD142, DPP4 and PDGFRA. ( C ) PDGFRA + DPP4+ double-positive cells from the stromal vascular fractions were cultured in adipogenic conditions for 12 days. The cells were stained with Oil Red O or collected for RNA analysis. The Oil Red O was extracted and quantified at 492 nM. Expression of Adipoq , Pparg and Prdm16 was determined by qPCR (n = 3; representative of 3 replicate experiments). Data are shown as mean ± SEM; (A and C) unpaired two-tailed Student's t test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The following antibodies were used: Perilipin1 (CST, 1:500), CD31 (R&D systems, 1:200), CD142 (Sino Biological, 1:50), MGP (Abcam, 1:2000), ICAM1 (Proteintech, 1:200), incubated at 4 °C overnight.

Techniques: Expressing, Knock-Out, Isolation, Cell Culture, Staining, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Two-step regulation by matrix Gla protein in brown adipose cell differentiation

doi: 10.1016/j.molmet.2024.101870

Figure Lengend Snippet: Single cell RNA-seq data analysis of GEO dataset GSE207707 . ( A ) t-Stochastic neighboring embedding (tSNE) plot of brown adipose stromal/stem cells (ASCs) and vascular cells. EC, endothelial cells; SMC, smooth muscle cells. ( B ) Scatter plot of Mgp expression in brown adipose ASCs and vascular cells. ( C ) Violin plots of the marker genes of brown adipose progenitor cells and pre-adipocytes. ( D ) Single-cell trajectory of brown adipose ASCs and the expression of marker genes along the trajectory. ( E ) Brown adipose ASCs ordered by pseudotime along the single-cell trajectory and gene expression dynamics of Mgp and Cd142/F3 in the ASC differentiation course.

Article Snippet: The following antibodies were used: Perilipin1 (CST, 1:500), CD31 (R&D systems, 1:200), CD142 (Sino Biological, 1:50), MGP (Abcam, 1:2000), ICAM1 (Proteintech, 1:200), incubated at 4 °C overnight.

Techniques: RNA Sequencing Assay, Expressing, Marker