Ex vivo flow-sorted calvarial osteoblasts from wildtype and Hrpt2 conditional KO neonatal mice. Mus musculus breed:C57BL/6

In order to determine how loss of Hrtp2 changes gene expression within mature osteoblasts and osteocytes, Ocn-CreTG/+;Hrtp2flox/flox mice were crossed to mT/mG Cre-reporter mice. The mT/mG transgene is a double-fluorescent Cre reporter that expresses membrane-targeted tandem dimer Tomato (mT) prior to Cre-mediated excision and expresses membrane-targeted green fluorescent protein (mG) after excision. Using this approach, matings between transgenic homozygotes to produce litters of pups with GFP-positive wild-type and GFP-positive, Hrtp2-deficient osteoblasts. This enabled us to isolate highly enriched populations of wild-type (Ocn-CreTG/TG;mTmGKnock-in) and Ocn-CreTG/TG;Hrtp2flox/flox;mTmGknock-in calvarial osteoblasts via flow cytometry. RNA was isolated from flow-sorted osteoblasts followed by RNA sequence analysis, which revealed that Hrtp2-deficient osteoblasts have increased expression of genes associated with cell morphogenesis, ossification, and skeletal development.