Figure 1.

N1 addition to MNLC-MG co-cultures influences whole culture and MG morphology after 7 days. (a) Timeline schematic showing differentiation of the MNLCs, MG introduction, N1 addition and experimental observation points. (b) Prestoblue metabolism of the N1or PBS treated MNLC-MG co-cultures shown as a percentage of the MNLC alone responses (dotted line) at 24 hours and 7 days. n = 4. (c) Low magnification (10×), live cell MitoTracker-green staining after 7 days with or without N1. Scale bar = 50 µm. (d) Quantification of the surface area covered by the cultures after 7 days co-culture. n = 3. (e) Live imaging of MG morphology at 24 hours and 7 days after addition of N1. Scale bar = 20 µm. (f) Count of the number of microglia displaying ramified morphology under control and N1 treated conditions after 7 days. Cell sketches shown above the graph represent examples of MG classified as non-ramified vs ramified. n = 3. (g) Count of the number of microglia associated with MNLC tissue after 7 days. n = 3. (h) MG morphology and association with different cell types following fixing and staining. Scale bar = 15 µm. Further NF-L staining and no primary background control staining is shown in Sup Fig. 1. Graphs show the mean and s.e.m. of ‘n’ independent repeats. *p < 0.05, ***p < 0.001.