PMC:1942070 / 3700-4529 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/1942070","sourcedb":"PMC","sourceid":"1942070","source_url":"http://www.ncbi.nlm.nih.gov/pmc/1942070","text":"2 Materials and methods\n\n2.1 Cell culture, transient transfections and cell stimulation\nThe generation, culture and activation of PKD1−/−, PKD3−/− and PKD1/3−/− knockout DT40 B cell lines have been described previously [1]. Cells were lysed and protein extracts were analysed in Western blotting experiments as previously described [1]. Chloramphenicol acetyl transferase assays have been described previously [29].\n\n2.2 sIgM staining\nDT40 B cells (2 × 106 cells per point) were resuspended in 200 μl buffer (RPMI 1640 media, 1% foetal calf serum) containing anti-chicken M1 monoclonal antibody conjugated to FITC for 20 min on ice. The cells were washed twice and fluorescent intensity was analysed by flow cytometry.\nAll results shown are representative of at two to four independent experiments unless otherwise 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