Journal: Carcinogenesis

Article Title: The lung-enriched p53 mutants V157F and R158L/P regulate a gain of function transcriptome in lung cancer

doi: 10.1093/carcin/bgz087

Figure Lengend Snippet: uPA protein expression is mutp53-dependent. Protein expression of uPA encoded by the RNA-seq identified gene PLAU, was assessed in p53-depleted and control conditions. (A) Western blot of A549, H2087, H441, H2110 and H2009 cells transfected with negative control or siRNA directed against p53 (25 nM). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is shown as a loading control. (B) qRT–PCR was performed to evaluate mRNA expression of PLAU after depletion of p53 by siRNA. mRNA expression values are normalized to GAPDH and siCtrl condition within each cell line. (C) Protein and conditioned media were harvested from cells after overnight incubation in reduced-serum conditions. Nitrocellulose membranes were stained with Ponceau S dye to visualize protein bands for loading control. Protein expression of pro-uPA (49 kDa) and its cleaved and activated forms B-chain (33 kDa) and A-chain (19 kDa) are shown in cell lysates and conditioned media. (D) Protein and E. mRNA expression of uPA (PLAU) following depletion of V157F mutp53 by siRNA.

Article Snippet: The following antibodies were used to detect protein expression by western blot: p53 (DO-1, sc-126; Santa Cruz), p21 (C-19, sc-397, Santa Cruz) and uPA (AF1310, R&D systems).

Techniques: Expressing, RNA Sequencing, Control, Western Blot, Transfection, Negative Control, Quantitative RT-PCR, Incubation, Staining

Journal: iScience

Article Title: Revealing the role of cancer-associated fibroblast senescence in prognosis and immune landscape in pancreatic cancer

doi: 10.1016/j.isci.2024.111612

Figure Lengend Snippet: PLAU derived from SCAFs promotes PDAC progression in vitro and in vivo (A) Relative mRNA expression of genes in human CAFs treated with 400 μM H 2 O 2 was measured by qRT-PCR. (B) The mRNA expression levels of genes in various cell types in the TME of PDAC. (C) The relationship between PLAU expression and UCell senescence score in CAFs. (D) PLAU knockdown in human SCAFs was verified by ELISA. (E) The migration and invasion abilities of the indicated cells were assessed by Transwell assay. Scale bar, 20 μm. (F) PLAU overexpression in human CAFs was verified by ELISA. (G) The migration and invasion abilities of the indicated cells were assessed by Transwell assay. Scale bar, 20 μm. (H) The flow of the experimental design. (I) Bioluminescence images showing orthotopically transplanted PDAC tumors. (J) Photographs and volumes of tumors. (K and L) Representative flow cytometry images and statistical analysis of CD3 + (K) and CD8 + T cells (L) in tumors in different groups. (M and N) Representative flow cytometry images and statistical analysis of MDSCs (M) and M2 macrophage (N) in tumors in different groups. Data are represented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Human uPA/Urokinase/PLAU ELISA Kit , BOSTER , Cat#EK0535.

Techniques: Derivative Assay, In Vitro, In Vivo, Expressing, Quantitative RT-PCR, Knockdown, Enzyme-linked Immunosorbent Assay, Migration, Transwell Assay, Over Expression, Flow Cytometry

Journal: iScience

Article Title: Revealing the role of cancer-associated fibroblast senescence in prognosis and immune landscape in pancreatic cancer

doi: 10.1016/j.isci.2024.111612

Figure Lengend Snippet:

Article Snippet: Human uPA/Urokinase/PLAU ELISA Kit , BOSTER , Cat#EK0535.

Techniques: Purification, Blocking Assay, Virus, Plasmid Preparation, Recombinant, Saline, Staining, Marker, Bicinchoninic Acid Protein Assay, CCK-8 Assay, Infection, Enzyme-linked Immunosorbent Assay, Gene Expression, Microarray, Expressing, Software