Fig. 6

Mixture effects on synaptogenesis. hiPSC-derived NSCs were differentiated for 7 DIV, and then treated for either 3 days (a and c) or 14 days (b and d) with single chemicals (BPA, CPF, Lead, Methyl-Hg, PCB138 and VA) and three types of mixtures: (i) a mixture with the 3 similar MoA chemicals (labelled ‘3-Sim’), (ii) a mixture with the 3 dissimilar MoA chemicals (labelled ‘3-Diss’), and (iii) a mixture with all 6 chemicals (labelled ‘All’). (a, b) Graphs reporting total levels of PSD95 (red), total levels of SYP (green) and number of overlapping SYP/PSD95 spots (synapses, yellow) analyzed upon treatment with LOAEC-syn concentrations. (c, d) Representative immunocytochemical image (at 10x magnification, with 40x magnifications insets) of cells treated with mixtures at different LOAEC-syn concentrations (see Table 3) for either 3 days (c) or 14 days (d) and stained for PSD95 (red) and SYP (green). 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