Figure 4.

Expression of Notch2 target genes and mitogen-activated protein kinase and phosphatidylinositol 3 kinase (PI3K)/Akt signaling pathway elements in Notch2HCS and littermate control mice. A: Total RNA from CD19⁺ cells was extracted and gene expression determined by quantitative RT-PCR. B and C: Western blot analysis of total cell lysates (100 μg) from spleen cells was fractionated by SDS-PAGE, and blots were probed in B with anti-Notch2 antibodies to detect the Notch2 intracellular domain (N2ICD) and in C with anti–phospho-Akt (pAkt) Ser473 and anti–phospho-Jun N-terminal kinase (pJNK) antibodies. Blots were stripped and reprobed with anti-Akt, anti-JNK, or anti–β-actin antibodies to verify protein loading. The ratios of pAkt/Akt to pJNK/JNK were quantified by Odyssey Infrared Imaging Systems software version 2.1 (LI-COR Biosciences, Lincoln, NE) and are shown under each blot. Molecular weight (M_r) ladder is shown along the left. Data are expressed as mean (95% CI) copy numbers of Notch2^6955T>C mutant, Hes5, and Hes1, corrected for Rpl38 (A). n = 4 biological replicates (A). ^∗P < 0.05 between Notch2HCS mutant and wild-type control cells (A).