Fig. 6.

Mixture effects on percentages of neurons and astrocytes. NSCs were differentiated for 7 DIV, and then treated for 14d with single chemicals and three types of mixtures as described in Fig. 3 legend. (A, B) Graphs report percentage of β-III-tubulin⁺ (A) and GFAP⁺ cells (B) analysed upon 14d treatment with LOAEC-neu concentrations (see Table 2). (C) Representative immunocytochemical images (at 10x magnification) of cells treated with neurite outgrowth-related mixtures (at LOAEC/2-neu concentrations) and stained for β-III-tubulin (red) and GFAP (green). All samples were normalised to medium containing solvent only (0.1 % DMSO, Ctr). Data are represented as mean ± S.E.M. of 3-4 biological replicates. (D) Heatmaps comparing the effects elicited by individual chemicals and mixtures, and showing the contribution of each chemical to overall mixture effect (i.e., dark green: high correlation, same effect directionality; dark yellow: low correlation, opposite effect directionality). (E) Correlation plots comparing mixture effects (size and colour of the circles corresponds to the level of correlation between pair-wise mixtures: i.e., the bigger and darker green, the higher the level of correlation). D and E were calculated according to Pearson and the correlation matrices were calculated considering the response upon exposure to LOAEC/2-, LOAEC/4- and LOAEC/8-neu and effect trend/directionality.