Characterization of novel pre-B-cell lines generated from BASH-deficient mice. (A) Flow cytometric analysis of the receptor expression on the indicated cell lines. Cells were stained for pre-BCR (left), κ chain (middle), and CD19 (right). (B) The indicated cell lines were stimulated (+) or not (-) with goat anti-mouse μH chain antibody, F(ab′)2 fragment (anti-μ, 10 μg/mL) for 1 minute. Cell lysates were subjected to immunoprecipitation (IP) with anti-Btk or anti-PLCγ2 antibody, and the precipitates were subjected to Western blot analysis using antiphosphotyrosine antibody (pY). The same filters were reprobed with anti-Btk or anti-PLCγ2 antibody as indicated to confirm the protein load. (C) The indicated cell lines were stimulated with anti-μ (10 μg/mL) for 0, 3, and 10 minutes. Cell lysates were subjected to Western blot analysis using antiphospho-ERK2 (pERK2) and antiphospho-JNK (pJNK) antibodies. The same filters were reprobed with anti-ERK2 and anti-JNK antibodies.
