Fig. 6
Vhlh is deleted in granulocytes of HOXB7-Cre; Vhlhfl/fl mice. a Isolation of (1) non-myeloid cells (CD11b-), (2) other myeloid cells (CD11b + SSCA low/medium), and (3) granulocytes (CD11b + SSCA high) from HOXB7-Cre; Vhlhfl/fl mice by FACS. Shown are representative FACS-plots before and after sorting. The three populations are indicated in each panel. b PCR analysis of Vhlh alleles in genomic DNAs isolated from the above cell populations. Recombined Vhlh allele (Vhlh del) is consistently observed in (3) granulocyte-enriched fraction (~95 % CD11b+, 70 % CD11b + SSCA high). As control, PCR was performed without template (no template control) or with genomic DNA from the liver of a HOXB7-Cre; Vhlhfl/+ mouse (positive control containing Vhlh-floxed and wild-type cells) or a HOXB7-Cre; Vhlhfl/fl kidney (positive control containing Vhlh-del cells). c Consistent with Vhlh inactivation, the mRNA of Proline-hydroxylase 3 (Phd3), a Hif-2α-responsive gene, is up-regulated in the granulocyte fraction, as assayed by qPCR