Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: PPY-Induced iCAFs Cultivate an Immunosuppressive Microenvironment in Pancreatic Cancer.

doi: 10.1002/advs.202413432

Figure Lengend Snippet: Figure 1. Screening and initial validation of cancer cell-secreted proteins capable of significantly inducing iCAF phenotype. A) The t-distributed stochastic neighbor embedding (t-SNE) plot of the 92,222 cells in the single-cell sequencing profile revealed distinct cell types observed in PDAC. B) The t-SNE plot exhibited diverse subtypes of fibroblasts observed in PDAC. C) The top 10 up-regulated and down-regulated expressed marker genes of each CAF subgroup. D) The expression levels of myCAF markers (ACTA2, COL1A1, COL11A1, MMP11), iCAF markers (CXCL12, IL-6, CCL2, CXCL2), and apCAF markers (HLA-DRA, HLA-DRB1) in different fibroblast subsets. E) Pathway activities scored by GSVA between different fibroblast subsets. F) The volcano

Article Snippet: ELISA assays used were Mouse CXCL12 ELISA kit (KE10049, Proteintech), Mouse IL-6 ELISA kit (EK206HS, MULTI SCIENCES), Mouse CCL2 ELISA kit (EK287, MULTI SCIENCES), Human CXCL12 ELISA kit (EK1119, MULTI SCIENCES), Human CCL2 ELISA kit (EK187, MULTI SCIENCES), and Human IL-6 ELISA kit (EK106, MULTI SCIENCES).

Techniques: Biomarker Discovery, Sequencing, Marker, Expressing

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: PPY-Induced iCAFs Cultivate an Immunosuppressive Microenvironment in Pancreatic Cancer.

doi: 10.1002/advs.202413432

Figure Lengend Snippet: Figure 2. PPY significantly induces the iCAF phenotype in PDAC CAFs both in vitro and in vivo. A–C) After treating CAFs derived from human PDAC tissues for 12 h, qRT-PCR analysis was performed to assess their alterations in the expression of iCAF markers (CXCL12 (A), IL-6 (B), CXCL12 (C)). D–F) qRT-PCR analysis of iCAF markers (CXCL12 (D), IL-6 (E), CXCL12 (F)) after treating the human CAFs for 24 h. G–I) qRT-PCR analysis of iCAF markers (CXCL12 (G), IL-6 (H), CXCL12 (I)) after treating the human CAFs for 36 h. J) qRT-PCR analysis of the expression levels of myCAF markers (ACTA2 and CTGF) after treating the human CAFs with PPY proteins (40ng/ml) for 24 h. K) Flow cytometry analysis was performed to evaluate the

Article Snippet: ELISA assays used were Mouse CXCL12 ELISA kit (KE10049, Proteintech), Mouse IL-6 ELISA kit (EK206HS, MULTI SCIENCES), Mouse CCL2 ELISA kit (EK287, MULTI SCIENCES), Human CXCL12 ELISA kit (EK1119, MULTI SCIENCES), Human CCL2 ELISA kit (EK187, MULTI SCIENCES), and Human IL-6 ELISA kit (EK106, MULTI SCIENCES).

Techniques: In Vitro, In Vivo, Derivative Assay, Quantitative RT-PCR, Expressing, Flow Cytometry

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: PPY-Induced iCAFs Cultivate an Immunosuppressive Microenvironment in Pancreatic Cancer.

doi: 10.1002/advs.202413432

Figure Lengend Snippet: Figure 7. The inhibition of EGFR expression in CAFs impeded the induction of iCAFs by PPY. A) qRT-PCR (A) and B) ELISA analyses of the expression levels of IL-6, CCL2, and CXCL12 in human EGFR-knockdown CAFs treated with PPY proteins. C) The efficiency of EGFR knockdown in KPC CAFs was examined by qRT-PCR. D,E) qRT-PCR (D) and ELISA (E) analyses of the expression levels of IL-6, CCL2, and CXCL12 in murine EGFR knockdown CAFs treated with PPY proteins. F) Flow cytometry analysis was performed to evaluate the populations of iCAFs, myCAFs, and apCAFs in murine EGFR- knockdown CAFs treated with PPY proteins. G,H) The IVIS image (G) and gross image (H) of tumors in model mice (n = 7), that was constructed

Article Snippet: ELISA assays used were Mouse CXCL12 ELISA kit (KE10049, Proteintech), Mouse IL-6 ELISA kit (EK206HS, MULTI SCIENCES), Mouse CCL2 ELISA kit (EK287, MULTI SCIENCES), Human CXCL12 ELISA kit (EK1119, MULTI SCIENCES), Human CCL2 ELISA kit (EK187, MULTI SCIENCES), and Human IL-6 ELISA kit (EK106, MULTI SCIENCES).

Techniques: Inhibition, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Knockdown, Flow Cytometry, Construct

Journal: Neuroscience

Article Title: Effects of crenolanib, a non-selective inhibitor of PDGFR, in a mouse model of transient middle cerebral artery occlusion

doi: 10.1016/j.neuroscience.2017.09.025

Figure Lengend Snippet: (A–C) Immunofluorescence staining of proliferative vessels (CD31+/BrdU+) in the peri-infarct region on day 7 after MCAO. CD31: green, BrdU: red. Images are shown at 200× magnification; scale bar=50 μm. (D) Quantification showed that crenolanib treatment on days 1–3 significantly decreased the number of proliferative vessels in the peri-infarct area on day 7 after stroke. *p<0.05 vs. MCAO+vehicle group; n=8/group. (E) ELISA analysis of ischemic border on day 7 after MCAO showed that crenolanib treatment on days 1–3 decreased the average level of VEGF in MCAO mice, but the change was not statistically significant. n=8/group. (F–G) Immunofluorescence staining of SDF-1 and MCP-1 in the peri-infarct region on day 7 after MCAO. The white lines show the brain boundaries. Images are shown at 100× magnification; scale bar=25 μm. (H–I) ELISA analysis of SDF-1 and MCP-1 on day 7 after MCAO showed that levels of SDF-1 and MCP-1 in the ischemic border were lower in MCAO mice treated with crenolanib (on days 1–3) than in MCAO mice treated with vehicle. *p<0.05 vs. MCAO+vehicle group; n=8/group.

Article Snippet: Based on our established protocol ( Zhao et al., 2015 , Han et al., 2016 , Wang et al., 2017 ), brain sections (20 μm) were processed with Nissl staining to quantify infarct volume or stained with antibodies against PDGFRβ (1:300, Abcam, Cambridge, MA, USA), GFAP (1:300, Santa Cruz Biotechnology, Dallas, TX, USA), doublecortin (DCX; 1:250, Santa Cruz Biotechnology), cleaved-caspase 3 (cCasp3; 1:500, Millipore, Billerica, MA, USA), CD31 (1:100, Abcam), BrdU (1:250, Abcam), Lectin fluorescein lycopersicon esculentum (tomato) (Lectin, 1:1000; Vector Laboratories, Burlingame, CA), SDF-1 (1:50, Proteintech, Sanying Biotechnology, Wuhan, China), MCP-1 (Proteintech), CD68 (1:500, Santa Cruz Biotechnology), occludin (1:250, Proteintech), and albumin (1:500, Santa Cruz Biotechnology).

Techniques: Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay