Journal: The Journal of Biological Chemistry

Article Title: Dynamic Heterogeneity of Brachyury in Mouse Epiblast Stem Cells Mediates Distinct Response to Extrinsic Bone Morphogenetic Protein (BMP) Signaling *

doi: 10.1074/jbc.M115.705418

Figure Lengend Snippet: T is critical for EMT process during EpiSC mesendoderm differentiation. A, expression levels of T and EMT-related genes in control and T-knockdown EpiSCs in unbiased differentiation at day 4 were determined by qPCR, including epithelial marker gene E-cadherin, mesenchymal marker genes N-cadherin and fibronectin, and transcriptional factors Snail and Zeb2. B, Western blotting analysis of T, E-cadherin, N-cadherin, fibronectin, Snail, phospho-ERK (p-Erk), and ERK in control and T-knockdown EpiSCs during 3 days of unbiased differentiation process. D, day; C, control; KD, knockdown. C, expression levels of mesoderm marker genes Flk1 and Mixl1 and endoderm marker genes Gata6 and Sox17 in control and T-knockdown EpiSCs during 4 days of unbiased differentiation were determined by qPCR. D, immunocytochemical assays (a) and statistical analysis (b) of T, Flk1, and Gata6, in day 3 EBs described in C. Scale bar, 75 μm. Error bars, S.E.

Article Snippet: The following antibodies were used: anti-phosphorylated Smad1/5/8 (1:2000; Cell Signaling Technology), anti-Smad1 (1:2000; Cell Signaling Technology), anti-phosphorylated ERK1/2 (1:1000; Cell Signaling Technology), anti-ERK1/2 (1:2000; Santa Cruz Biotechnology), anti-T (1:1000; R&D Systems), anti-β-actin (1:7000; Abgene), anti-Snail (1:1000; Abcam), anti-Id (1:1000; Abcam), anti-N-cadherin (1:2000; Invitrogen), anti-E-cadherin (1:1000; Santa Cruz Biotechnology), anti-Fibronectin (1:1000; Abcam), and anti-β-Catenin (1:1000; Santa Cruz Biotechnology).

Techniques: Expressing, Marker, Western Blot

Journal: Nephrology Dialysis Transplantation

Article Title: The Raf kinase inhibitor PLX5568 slows cyst proliferation in rat polycystic kidney disease but promotes renal and hepatic fibrosis

doi: 10.1093/ndt/gfr432

Figure Lengend Snippet: PLX5568 increased renal and hepatic fibrosis in cy/+ rats. (A) Evaluation of kidney sections stained positive with picrosirius red of cy/+ rats treated either with vehicle or PLX5568 standardized for the difference of cystic area (number of kidneys: Ctrl: n = 8, PLX: n = 7, *P = 0.04). Area of kidney sections of control and PLX5568-treated animals stained positive for fibronectin (B), matrix metalloproteinase-2 (MMP2) (C) and plasminogen activator inhibitor-1 (PAI 1) (D) standardized for the difference of cystic area (number of kidneys: Ctrl: n = 8, PLX: n = 7, *P = 0.016 for fibronectin and *P = 0.0006 for MMP2 and PAI 1). (E) Number of cells stained positive for S100 calcium-binding protein A4 (FSP1) per tubule (number of kidneys: Ctrl: n = 8, PLX: n = 7, *P = 0.01). (F) Evaluation of liver sections stained positive with picrosirius red of cy/+ rats treated either with vehicle or PLX5568 (number of livers: Ctrl: n = 8, PLX: n = 7, *P = 0.04).

Article Snippet: For detection of fibrosis, the following antibodies were used: mouse anti-fibronectin (1:500; Chemicon, Billerica, MA), goat anti-MMP2 (1:50; Santa Cruz Biotechnology Inc., Santa Cruz, CA), goat anti-PAI 1 (1:50; american diagnostica GmbH, Pfungstadt, Germany), rabbit anti-FSP1 (1:100; Abcam, Cambridge, UK).

Techniques: Staining, Binding Assay