Journal: Cell death discovery

Article Title: Serpine1 mRNA confers mesenchymal characteristics to the cell and promotes CD8+ T cells exclusion from colon adenocarcinomas.

doi: 10.1038/s41420-024-01886-8

Figure Lengend Snippet: Fig. 1 Argonaute iCLIP. A Location of AGO2-mRNA tags in the genome of control NMuMG cells and cells treated for 4, and 48 h with TGF-β. B Differential association of transcripts to AGO2 after 4 h of TGF-β treatment [Log2(Fold Change 4 h of TGF-β versus Control)], respect to transcript mean expression. The Serpine1 point was highlighted in red. C Distribution of the AGO2 34 crosslinking sites along the Serpine1 mRNA. 3´UTR, 5´UTR, coding region and exons of the Serpine1 mRNA are indicated.

Article Snippet: The following antibodies were used in our study: anti-TRA2B (Santa Cruz; sc166829), anti-PGM1 (Invitrogen; PA5-55008), anti-ADH7 (Invitrogen; PA597437), anti-mouse Serpine1 (R&D Systems; AF3828), anti-human Serpine1 (ThermoFisher, #66261-1-IG), anti-CD8A (Sigma-Aldrich, HPA037756), antiAGO2 (Wako Chemicals; clone 2D4, # 018-22021).

Techniques: Control, Expressing

Journal: Cell death discovery

Article Title: Serpine1 mRNA confers mesenchymal characteristics to the cell and promotes CD8+ T cells exclusion from colon adenocarcinomas.

doi: 10.1038/s41420-024-01886-8

Figure Lengend Snippet: Fig. 2 Serpine1 expression in TGF-β–treated NMuMG cells. Relative levels of mRNA Serpine1 (A) and protein SERPINE1 (B) in cells treated with TGF-β at the indicated times. A representative immunoblotting of SERPINE1 is shown. Quantification of Serpine1 and SERPINE1 in three independent experiments is shown. Error bars represent S.D. ***p < 0.001, **p < 0.01 by two-tailed Student´s t test. Protein-loading normalization was performed by measuring total protein directly on the membrane using the criterion stain-free gel imaging system.

Article Snippet: The following antibodies were used in our study: anti-TRA2B (Santa Cruz; sc166829), anti-PGM1 (Invitrogen; PA5-55008), anti-ADH7 (Invitrogen; PA597437), anti-mouse Serpine1 (R&D Systems; AF3828), anti-human Serpine1 (ThermoFisher, #66261-1-IG), anti-CD8A (Sigma-Aldrich, HPA037756), antiAGO2 (Wako Chemicals; clone 2D4, # 018-22021).

Techniques: Expressing, Western Blot, Two Tailed Test, Membrane, Staining, Imaging

Journal: Cell death discovery

Article Title: Serpine1 mRNA confers mesenchymal characteristics to the cell and promotes CD8+ T cells exclusion from colon adenocarcinomas.

doi: 10.1038/s41420-024-01886-8

Figure Lengend Snippet: Fig. 6 SERPINE1 mRNA expression is associated with the downregulation of genes involved in different pathways that regulate the innate immune response and decrease infiltrated lymphocytes in human colon adenocarcinomas. A, B Expression of SERPINE1 mRNA and SERPINE1 protein in COAD samples and adjacent normal samples from TCGA and CPTAC databases respectively. C The Spearman correlation coefficients in the analysis of the expression levels of SERPINE1 mRNA and the infiltration levels of CD8+ T cells, CD4 + T cells, B cells, macrophages and neutrophils in patients with COAD (n = 458). D, E Linear correlation analysis between SERPINE1 and CD8A mRNAs and SERPINE1 and CD3A mRNAs, respectively, in COAD surgical samples. F Representative images of COAD samples showing low, medium, and high signal for SERPINE1 mRNA, detected by in situ hybridization (ISH) and the corresponding immunofluorescence signal (IFC) using anti- CD8A antibody (red) and DAPI (blue).

Article Snippet: The following antibodies were used in our study: anti-TRA2B (Santa Cruz; sc166829), anti-PGM1 (Invitrogen; PA5-55008), anti-ADH7 (Invitrogen; PA597437), anti-mouse Serpine1 (R&D Systems; AF3828), anti-human Serpine1 (ThermoFisher, #66261-1-IG), anti-CD8A (Sigma-Aldrich, HPA037756), antiAGO2 (Wako Chemicals; clone 2D4, # 018-22021).

Techniques: Expressing, In Situ Hybridization