Gene expression profile of spleen B cells from Dock10 knockout mice and its regulation by IL-4. Mus musculus

The goals of this study were to identify alterations in the gene expression profile (GEP) of spleen B cells purified from Dock10 knockout (ko) mice [C57BL/6 Dock10tm1a(EUCOMM)Hmgu/Ieg, European Mouse Mutant Archive] by comparison with the GEP of spleen B cells purified from C57BL/6 wild-type (wt) mice, and to identify alterations in the GEP of spleen B cells of the Dock10 ko mice in response to culture with IL-4 during 18 hours, by comparison with the response of the wt mice. Overall design: Spleen B cells from 10 C57BL/6 wt mice and 10 C57BL/6 Dock10 ko mice of 12 weeks of age were purified by negative selection using the Miltenyi Pan B Cell Isolation Kit, mouse. Five of the wt samples and 5 of the ko samples, those numbered from 6 to 10 were cultured with nothing (Ctrl) or with mouse recombinant interleukin 4 (IL-4) during 18 hours. Equimolar RNA amounts from 5 of the same ko samples before culture (Pre), those numbered 6 to 10, were pooled, labelled with cy3, and used as reference sample. Each of the wt samples and the ko samples, including Pre, Ctrl, and IL-4, was labelled with cy5 and used as a test sample. The reference sample and the test samples were hybridized together onto Agilent Mouse GE 4x44K v2 Microarrays (technology 26655), scanned in an Agilent G2565CA DNA Microarray Scanner using the Agilent Scan Control software, processed with the Agilent Feature Extraction software, and analyzed using the GeneSpring GX software.