Supplementary Figure 4: Extreme limiting dilution assay (ELDA) of BTICs in absence or presence of media conditioned by human microglia and monocytes.
From: Therapeutic activation of macrophages and microglia to suppress brain tumor-initiating cells
Extreme limiting dilution assay (ELDA) was carried out as described previously25 (a,d). Briefly, BTIC cells (25EF and 48EF) were plated in decreasing numbers from 200 cells/well to 1 cell/well in 200μl. Cultures were maintained until day 14, when the number of wells containing spheres for each cell plating density (number of positive cultures) was recorded, calculated and plotted using online ELDA analysis program25 (http://bioinf.wehi.edu.au/software/elda). ELDA analysis for 25EF (b,c) and 48EF lines (e,f). ELDA experiment shows zero sphere formation with MCM or MonoCM in all wells including single cell wells compared to control (a and d), and ELDA indicates significant differences in stem cell frequencies between the groups (c and f). n=4 for all groups; Error bars, s.d.
