Fig. 6

Increased expression of Satb2 in the cortex of alcohol-exposed animals does not correlate with alterations in gene expression or increased H3K9me2. A Using RT-qPCR, we assayed levels of transcripts encoding the chromatin looping proteins Satb1 and Satb2 between control, alcohol-exposed but normal, and alcohol-exposed animals exhibiting structural defects. B Western blot analysis of SATB2 protein levels within the fetal cortex across treatment groups, with samples normalized to TUBULIN. C RT-qPCR analysis of Satb1 and Satb2 in the cerebellum and heart. D Using samples derived from the fetal cortex and ChIP-qPCR, we assayed the Satb1 and Satb2 promoters for the enrichment of H3K9me2. E As a positive control, we used ChIP-qPCR to determine the enrichment of H3K9me2 (left) and SATB2 (right) at the Fezf2 upstream enhancer, a known SATB2 binding site [53]. F Using ChIP-qPCR, we determined SATB2 enrichment at the indicated transposable elements. For the cortex, gene expression was normalized to transcripts encoding Gapdh and Ywhaz, while in the cerebellum and heart, expression was normalized to the transcripts encoding H2Afz and Ywhaz (n = 8 Control, 9 Unaffected, 7 Affected). Error bars represent SEM, *P < 0.05, **P < 0.01