Article Title: Pancreatic islets communicate with lymphoid tissues via exocytosis of insulin peptides
Figure Lengend Snippet: Characterization of circulating B:9-23 and its localization into lymphoid organs a , Unmodified synthetic B:9-23 (3 pmoles) was spiked into 1 ml PBS, purified by C18 tips, lyophilized, and analyzed by nLC-MS/MS. The data show the appearance of unmodified B:9-23 (left) together with oxidation of the cysteine on the 19 th position to cysteic acid (right). b , c , Alexa Fluor 488-conjugated B:9-23 peptide (100 μg) was injected intravenously into 4-week old B6, B6g7 and NOD mice. 1h later, the spleens and thymi were harvested, digested by liberase and DNase, and the binding to splenic and thymic APCs was measured by flow cytometry. b, Representative FACS plots showing the binding of B:9-23 to splenic XCR1 + and Sirpα + DC subsets as well as the B cells (upper). The bar graph summarizes cumulative results from individual mice (each point) pooled from three independent experiments. ns, not significant; **, P
Article Snippet: The sample was passed through C18 zip-tips (Pierce) and peptides then were eluted in 0.1% formic acid/95% acetonitrile and then dried with a Speed-Vac.
Techniques: Purification, Mass Spectrometry, Injection, Mouse Assay, Binding Assay, Flow Cytometry, Cytometry, FACS