anti krt 19  (Novus Biologicals)

Journal: Nature Communications

Article Title: Patient-derived pancreas-on-a-chip to model cystic fibrosis-related disorders

doi: 10.1038/s41467-019-11178-w

Figure Lengend Snippet: Characterization of pancreatic ductal epithelial cells. ( a – f , l : Characterization of organoids). a Characterization of pancreatic ductal organoids using epithelial cell markers, b cytokeratin 19 (KRT 19, c E-cadherin, e sodium transport channel (ENaC), and f ZO-1. d Hematoxylin and eosin (H&E) image shows the orientation of pancreatic ductal epithelial cells in spheroid of the organoid. ( g – k : Characterization of monolayer of pancreatic ductal epithelial cells (PDECs)). g Phase contrast and j H&E images show monolayer of PDECs formed from the organoids. Monolayer of PDECs showed positivity for tight junction h ZO-1, i F-actin and KRT 19, and k cystic fibrosis transmembrane conductance regulator (CFTR). l RNA-sequencing data was obtained from the pancreatic ductal organoids and verified the PDEC origin ( n = 4 sample preparation from the same patient). Data are mean ± SD. Scale bars: 10 µm ( k ), 20 µm ( b , c , e , f , h – j enlarge), 50 µm ( d , g , j ), and 500 µm ( a )

Article Snippet: Cells were then blocked using 1% goat serum (Sigma-Aldrich; #A8806-5G) for 1 h at room temperature and incubated with primary antibodies (diluted in antibody diluent (Invitrogen; #TA-125-ADQ) 1:100), anti-CFTR R1104 (Eric Sorscher lab, CF Center, University of Alabama, Birmingham, AL, USA [presently, Emory University, Atlanta, GA, USA]), anti-ZO-1 (BD Biosciences; #610967), anti-ENaC (Invitrogen; #PA1-920A), anti-KRT 19 (Invitrogen; #MA5-12663), anti-E cadherin (Cell Signaling Technology; #3195), anti-insulin (Cell Signaling; #C27C9), and anti-glucagon (Sigma; #G2654) overnight at 4 °C.

Techniques: RNA Sequencing Assay, Sample Prep