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Fig. 4. Notch1/RBPj{kappa} regulates Gata2 transcriptional activity. (A) Semiquantitative RT-PCR analysis of Gata2, Aml1 and Scl expression in 32D wild-type or N1IC-expressing cells. Representative PCR products after 35 and 40 cycles of two independent experiments are shown. Quantitated relative mRNA levels of Gata2, Aml1 and Scl are shown in the lower graph. (B) Chromatin immunoprecipitation with anti-N1 from 32D wild-type cells and 32D-N1IC cells. PCR detection of the Gata2, Aml1 and Scl, Hes1 and ß-globin promoters from the precipitates is shown. (C) Double in-situ hybridization with Gata2 and Notch1 on transverse section of wild-type E10.5 aortas. (D) Section of WISH that shows Hes1 expression in hematopoietic clusters budding from the aorta from E10.5. (E) Chromatin immunoprecipitation with anti-N1 ({alpha}-N1) from wild-type and RBPj{kappa} mutant whole E9.5 embryos. PCR detection of the Gata2 and Aml1 promoter is shown.