Journal: Experimental hematology

Article Title: Pharmacological inhibition of LSD1 and mTOR reduces mitochondrial retention and associated ROS levels in the red blood cells of sickle cell disease

doi: 10.1016/j.exphem.2017.02.003

Figure Lengend Snippet: Peripheral blood cells obtained from WT and SCD mice were stained with MitoTracker Green FM. Scale bar: 5μm B) Peripheral blood cells obtained from WT and SCD mice or SCD mice treated with RN-1 (2.5mg/kg), RN-1 (5mg/kg), or Sirolimus (5mg) were stained with TMRM and thiazole orange or CD71-APC conjugated antibody. RBCs were gated based on specific forward and side scatter properties and analyzed by flow cytometry. The bar graph shows the mean percentage of peripheral mitochondria retaining RBCs in WT vs. SCD vs. SCD treated with RN-1 (5mg), or Sirolimus (5mg).C) Representative flow cytometry plots showing the percent of mitochondria retaining RBCs located in the upper-left quadrant in wild type SCD and SCD mice treated with RN-1 (5mg) or SCD mice treated with Sirolimus (5mg). D) Peripheral blood from Control (HbAA) and SCD (HbSS) mice were incubated with the ROS probe CM-H2DCFDA after staining with TMRM and CD71-APC. The bar graph shows the mean percent of RBCs with high ROS in WT vs. SCD vs. SCD treated with RN-1 (5mg), or Sirolimus (5mg). E) Representative flow cytometry plots showing the percent of cells in the RBC fraction with ROS located in the upper-left quadrant in wildtype, SCD, and SCD mice treated with RN-1 (5mg) or Sirolimus (5mg). F) Pearson correlation plot showing the correlation between mitochondria and ROS levels in RBC of control and treated SCD mice.

Article Snippet: SCD (HbSS) mice also known as the Townes model mice (B6; 129-Hba tm1 (HBA) Tow Hbb tm2 (HBG1, HBB*) Tow /Hbb tm3 (HBG1, HBB) Tow /J and wild type (HbAA) mice (B6; 129- Hba tm1 (HBA) Tow , Hbb tm3 (HBG1, HBB) Tow ) were purchased from the Jackson Laboratories (Bar Harbor, ME).

Techniques: Staining, Flow Cytometry, Control, Incubation