Supplementary Figure 6: Cell cycle analyses by flow cytometry confirm the reduction of proliferation of BTICs but not their differentiated cells in response to microglia-derived factors.

The 25EF BTIC line was grown in BTIC medium or was differentiated by culture for 5d in 1% FBS medium. Cells were then exposed to MCM or AmpB-treated microglia conditioned medium (AmpB/MCM) for 72h and subjected to propidium iodide flow cytometry for DNA content. a) Representative panels show 32% of control BTIC cells were in the S-phase of the cell cycle, and this was reduced in BTICs exposed to MCM, and further attenuated in BTICs exposed to AmpB/MCM. In contrast, in cells differentiated by 1%FBS, there was no impact of MCM or AmpB/MCM. These results were reproduced across 3 sets of cultures (b). ^*p<0.05, ^***p<0.001 compared to respective controls (1 way ANOVA). (c) Finally, AmpB, MCM or monoCM from 2 subjects, and those collected from AmpB-treated cultures, were without effects on growth of U251 or U87 differentiated glioma lines. n = 4 for all groups; Error bars, s.d.