Fig. 5: DIPG cells are protected from chemo- and targeted- therapies by the presence of stromal cell types.

A Representative images demonstrating response to TMZ chemotherapy (400 μm) in GFP expressing SU-DIPG-6 tumor cells (T-6) alone and in 3D co-culture with microglia expressing tagRFP) and endothelial cells with no fluorescent marker (TEM-6) at 72 h post treatment in flat-bottom 96-well plates. B % tumor cells present over time for GFP-expressing DIPG cells alone (green) and in 3D co-culture (orange) after treatment with variable concentration of TMZ (0–400 μm). C Quantification of cellular growth in U-bottom 96-well plates and D corresponding representative IncuCyte images (T-6, T-13 and TEM-6, TEM-13) demonstrate response to variable concentration of Marizomib (0–32 nM) and Panobinostat (0–32 nM) from 1–4 days. Here the tumor cells express GFP fluorescence (Green) and microglia express tagRFP (red) and endothelial cells do not express any fluorescent protein. Response of SU-DIPG-6 and SU-DIPG-13 tumor cells alone is identified as T-6 and T-13, respectively (green solid and dotted line). Response of GFP-labeled tumor cells in TEM-6 and TEM-13 co-culture with microglia and endothelial cells is denoted as T-TEM-6 and T-TEM-13 (blue solid and dotted line). Response of microglia (quantified via tagRFP label) in TEM co-cultures is indicated as M-TEM-6 and M-TEM-13 (Grey dotted and solid line). In all cases, values of integrated green- or red- fluorescence intensity per well (from GFP and tagRFP expression) were averaged across 4 well replicates per experiment.