80% of either MSC conditioned medium (MSC-CM) or plain MEM alpha was added to respective treatment wells, and L-NAME was kept in the medium.
To assess the effect of a diffuse source of reelin, MAM treated slices were incubated in medium containing recombinant reelin (1 nM) or plain medium as a control.
The medium was subsequently replaced with 100 μl of glucose free KRB containing plant extract (62.5 - 500 μg/ml), glibenclamide (0.1 - 10 μM) or plain medium, and incubated at 37°C in a 5% CO2 for 1 h.
Based on these results, we prepared glioma CM, pre-incubating for 4 h the C6 cells with LI (or plain medium, in order to assess the effect of glioma under basal condition).
Carbohydrate assimilation tests for sucrose and raffinose were conducted in 96-well microplates, using 150-μl of a working solution composed of yeast nitrogen base (YNB) medium (Sigma-Aldrich, USA) with corresponding test sugar, or plain (only base medium), as described by Marimon et al. [ 18].
The orally administered stressors were as made up follows: 1xSYA containing 20mM paraquat (Sigma-Aldrich, Dorset, UK), 1xSYA containing 0.03% w/v DDT (Supelco Sigma-Aldrich, Dorset, UK), 1.5% agar medium containing 5%H2O22(Sigma-Aldrich, Dorset, UK) and 50g/l sucrose, or plain 1.5% agar medium for the starvation experiment.
Freshly obtained ex vivo PBMC were treated overnight in plain medium or with the following cytokines or combination, thereof: IL-2, IL-15, IL-4 or IL-7+IL-15.
In MAM treated slices incubated in plain medium or cocultured with control HEK cells, BrdU+ cells tend to scatter in all cortical layers (Figure 5A,B,J).
Slices were incubated in plain medium or in medium containing 1 nM of soluble recombinant NRG1 or a 30 nM concentration, which has been shown to promote migration along the radial glial scaffold in mice [23].
Blockade of either Dab1 or Pi3K resulted in continued severe disruption of vimentin+ processes in the presence of reelin whereas BLBP+ processes were unchanged compared to controls (slices cultured in plain medium or cocultured with control HEK cells) (Figure 4G H,J).
