Characterization of wild-type and STAT5^−/− C57Bl/6 mice.

<p>(A) Tail DNA from STAT5^+/+, STAT5^+/−, and STAT5^−/− C57Bl/6 mice was prepared and PCR were performed to amplify wild-type <i>stat5a</i> and <i>stat5b</i> DNA as well as the TK cassette. The primers are given in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000727#s4" target="_blank">Material and methods</a> section. (B) LSECs were isolated from wild-type (WT) and STAT5^−/− mice. Whole-cell extracts were prepared and analyzed for STAT5b and STAT5bΔN expression by immunoblot. We used an antibody (G-2) directed to an epitope mapping at the C-terminus of STAT5b. Equal loading of proteins on the gel was confirmed by probing the blots for β-actin. (C) Blood neutrophils were isolated from WT and STAT5^−/− mice. Whole-cell extracts were prepared and analyzed for STAT5b/STAT5bΔN (left panel) and STAT5a/STAT5aΔN (right panel) expression by immunoblot. For STAT5b/STAT5bΔN, we used the G-2 antibody. For STAT5a/STAT5aΔN, we used an antibody (L-20) raised against a peptide mapping at the C-terminus of STAT5a. Equal loading of proteins on the gel was confirmed by probing the blots for β-actin.