Fig. 7.

Mixture effects on synaptogenesis. 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-C) Graphs report total levels of SYP (A), PSD95 (B), and number of overlapping SYP/PSD95 puncta (synapses, C) analysed upon 14d treatment with all tested LOAEC-syn concentrations (see Table 2). (D) Representative immunocytochemical image (at 10x magnification, with 40x magnifications insets) of cells treated with mixtures at different LOAEC/2-syn concentrations and stained for SYP (green) and PSD95 (red). All samples were normalised to solvent control (0.1 % DMSO, Ctr) at the respective time point. Data are represented as mean ± S.E.M. of 3-4 biological replicates. (E) 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). (F) 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). E and F were calculated according to Pearson and the correlation matrices were calculated considering the response upon exposure to LOAEC/2-, LOAEC/4- and LOAEC/8-syn and effect trend/directionality.