PMC:4502372 / 13352-14259 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/4502372","sourcedb":"PMC","sourceid":"4502372","source_url":"https://www.ncbi.nlm.nih.gov/pmc/4502372","text":"Strains were pregrown in YPLactate to approximately 0.5 × 107 cells. Initial aliquots were taken and then galactose was added to each culture to a final concentration of 2%. Aliquots were taken at the indicated time points. Samples were spun at 3000 rpm and cell pellets washed twice with water before being frozen at −80°. The cell pellets were thawed and normalized to cell count before lysis. For analysis of Rad53 phosphorylation, protein lysates were prepared by trichloroacetic acid method as previously described (Foiani et al. 1994). For analysis of Tat2 protein levels, samples were prepared as previously described (Abe and Iida 2003). Before western analysis of Tat2, 50 µg of whole-cell extract was denatured in 5% SDS and 5% β-mercaptoethanol at 37° for 10 m. Western blots were probed with α-Flag (F3165; Sigma-Aldrich), α-Rad53 (provided by M. Foiani), and α-PGK (ab113687; Abcam) antibodies.","tracks":[{"project":"2_test","denotations":[{"id":"25943524-8289832-43313261","span":{"begin":535,"end":539},"obj":"8289832"},{"id":"25943524-14560004-43313262","span":{"begin":639,"end":643},"obj":"14560004"}],"attributes":[{"subj":"25943524-8289832-43313261","pred":"source","obj":"2_test"},{"subj":"25943524-14560004-43313262","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#a5ec93","default":true}]}]}}