Effects of FLVCR-deletion on intracellular heme and ROS in megakaryocytes. Bone marrow mononuclear cells from FLVCR-deleted and wild-type Cre mice were isolated and cultured in StemPro34 media with 100 ng/mL of recombinant human thrombopoietin for 5 days. (A) Intracellular heme levels in FLVCR-del and wild-type Cre cultured megakaryocytes. (B) Representative flow cytometry plot of megakaryocyte intracellular ROS. Intracellular ROS was determined by flow cytometry after staining with 5 μM CM-H2DCFDA for 30 minutes. Bold line represents wild-type Cre and the solid line represents FLVCR-deleted cells. (C) Real-time PCR for FLVCR, TBXAS1, NQO1, and HMOX1 gene expression in cultured megakaryocytes derived from FLVCR-deleted or wild-type Cre bone marrow mononuclear cells. Expression was normalized to β-actin. *P < .05.