Figure 2
From: Microenvironmental Hypoxia regulates FLT3 expression and biology in AML
FLT3 is down-regulated by hypoxia in Ba/F3 cells.
(A) OCI-AML3 and OCI-AML5 do not regulate FLT3 under hypoxia. Fold chance Optical Density (OD) was calculated compared to the respective value under 21% O2 and normalized to β-actin. (B) Ba/F3 cell lines stably transfected with FLT3 WT and FLT3 W51 (ITD) reliably down-regulate FLT3 at hypoxic conditions. (C) Down-regulation of FLT3 occurs in Ba/F3 W51 cells only below oxygen levels of 6% O2. (D) Ba/F3 W51 cells down-regulate FLT3 after 48 hours of culture under hypoxia. Fold change Optical Density (OD) was calculated for each time point compared to the respective value under 21% O2 and normalized to β-actin. (E) FLT3 down-regulation in Ba/F3 W51 cells is not a short-term stress hypoxic effect but is maintained for >9 days of culture under hypoxia. (F) Restoration of FLT3 expression after re-oxygenation of hypoxic Ba/F3 W51 cells with 21% O2. (G) Optical Density (OD) of FLT3 was calculated at different conditions compared to the respective value under 21% O2 at 72 hours and normalised to β-actin. (H) Significant decrease in proliferation in Ba/F3 W51 cells after 48 h and 72 h of culture at 1% O2. (I) No increased induction of cell death by hypoxia of 1% O2 in Ba/F3 W51 cells after 72 hours as compared to cells at 21% O2.
