Figure 6: Patient variants in hs576 (VE854) reduce enhancer activity in cranial nerves.

(A) The genomic region containing hs576 and SIM1. (B) The locations of two ASD patient variants from the HMCA and NIMH cohorts (ASD; red), two variants from one control individual (ctrl; gray), and four previously identified obesity-associated variants (blue) (Kim et al., 2014) are indicated. These variants are all located in the most conserved region of hs576 (core), which recapitulates most of the enhancer activity of the entire hs576 element (Kim et al., 2014). (C) Constructs containing hs576 without (n = 4) or with (n = 6) the two ASD patient variants upstream of a minimal promoter driving the lacZ gene were integrated into the safe-harbor H11 locus and analyzed for lacZ expression at E11.5 (Materials and Methods). Arrowheads indicate cranial nerves where the inclusion of the two ASD patient variants reduces enhancer activity. Representative embryos are shown (all embryos are in Fig. S17). Coordinates are in hg19.