se r d systems  (R&D Systems)

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: The Human PLAU ELISA Kit and Mouse PLAU ELISA kit were purchased from BOSTER (Wuhan, China) and Hengyuan Biological Technology (Shanghai, China).

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: The Human PLAU ELISA Kit and Mouse PLAU ELISA kit were purchased from BOSTER (Wuhan, China) and Hengyuan Biological Technology (Shanghai, China).

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

Journal: JCI insight

Article Title: Transcriptional heterogeneity of fibroblasts is a hallmark of the aging heart.

doi: 10.1172/jci.insight.131092

Figure Lengend Snippet: Figure 6. Age-dependent changes of fibroblast function on endothelial cells. (A and B) Experimental outline and tube formation assay of human umbilical vein endothelial cells (HUVECs) that were cultured in conditioned medium received from young (12 weeks) and aged (20 months) cardiac mouse fibroblasts. Accumulated tube length was measured in 5 randomly chosen microscopic fields with a computer-assisted microscope using Axiovision 4.5 (Zeiss) (scale bar: 100 μm; n = 4). (C) Capillary density of random healthy areas of young (12 weeks; n = 6) and old (n = 8) heart sections versus capillary density of fibrotic (n = 8) areas of aged hearts (20 months). Shown is the quantification of the Isolectin B4–positive area versus the total area (%). (D) Expression of Serpine1, Serpine2, and Efemp1 in fibroblasts displayed in log scale in trajectory plots. (E) Secretion of SerpinE1 in isolated young and aged cardiac fibroblasts cultured for 24 hours in serum-free medium. Data were derived using a mouse angiogenesis array (R&D Systems) of culture supernatants of isolated cardiac fibroblasts (n = 4). (F) Tube formation assay of HUVECs that were cultured in conditioned medium received from young (12 weeks) and aged (20 months) cardiac mouse fibroblasts. The aged phenotype was rescued by supplementing the aged fibroblast medium with 4 μg of an anti-Serpin antibody. Accumulated tube length was mea- sured in 5 randomly chosen microscopic fields with a computer-assisted microscope using Axiovision 4.5 (Zeiss) (scale bar: 200 μm; n = 4). (G) RNA expression

Article Snippet: For the recombinant serpin study, HUVECs were cultured in medium supplemented with 10 ng/mL serpin E1 (CSB-EP021081MO, Cusabio) or serpin E2 (2175-PI-010, R&D Systems) for 24 hours.

Techniques: Tube Formation Assay, Cell Culture, Microscopy, Expressing, Isolation, Derivative Assay, RNA Expression

Journal: JCI insight

Article Title: Transcriptional heterogeneity of fibroblasts is a hallmark of the aging heart.

doi: 10.1172/jci.insight.131092

Figure Lengend Snippet: Figure 6. Age-dependent changes of fibroblast function on endothelial cells. (A and B) Experimental outline and tube formation assay of human umbilical vein endothelial cells (HUVECs) that were cultured in conditioned medium received from young (12 weeks) and aged (20 months) cardiac mouse fibroblasts. Accumulated tube length was measured in 5 randomly chosen microscopic fields with a computer-assisted microscope using Axiovision 4.5 (Zeiss) (scale bar: 100 μm; n = 4). (C) Capillary density of random healthy areas of young (12 weeks; n = 6) and old (n = 8) heart sections versus capillary density of fibrotic (n = 8) areas of aged hearts (20 months). Shown is the quantification of the Isolectin B4–positive area versus the total area (%). (D) Expression of Serpine1, Serpine2, and Efemp1 in fibroblasts displayed in log scale in trajectory plots. (E) Secretion of SerpinE1 in isolated young and aged cardiac fibroblasts cultured for 24 hours in serum-free medium. Data were derived using a mouse angiogenesis array (R&D Systems) of culture supernatants of isolated cardiac fibroblasts (n = 4). (F) Tube formation assay of HUVECs that were cultured in conditioned medium received from young (12 weeks) and aged (20 months) cardiac mouse fibroblasts. The aged phenotype was rescued by supplementing the aged fibroblast medium with 4 μg of an anti-Serpin antibody. Accumulated tube length was mea- sured in 5 randomly chosen microscopic fields with a computer-assisted microscope using Axiovision 4.5 (Zeiss) (scale bar: 200 μm; n = 4). (G) RNA expression

Article Snippet: For the recombinant serpin study, HUVECs were cultured in medium supplemented with 10 ng/mL serpin E1 (CSB-EP021081MO, Cusabio) or serpin E2 (2175-PI-010, R&D Systems) for 24 hours.

Techniques: Tube Formation Assay, Cell Culture, Microscopy, Expressing, Isolation, Derivative Assay, RNA Expression