1. Identification of the Transcriptional Targets of FOXP2, a Gene Linked to Speech and Language, in Developing Human Brain 
Elizabeth Spiteri, Genevieve Konopka, Giovanni Coppola, Jamee Bomar, Michael Oldham, Jing Ou, Sonja C. Vernes, Simon E. Fisher, Bing Ren, Daniel H. Geschwind 
The American Journal of Human Genetics 
Volume 81, Issue 6, Pages (December 2007)
DOI: /522237
Copyright © 2007 The American Society of Human Genetics Terms and Conditions

2. Figure 1
FOXP2 antibody detection of human brain and lung FOXP2 expression. A, Peptide used as an antigen to create the FOXP2 antibody aligned against the amino acid sequence of FOXP2 and family members FOXP1 and FOXP4. Protein from MRC5 cells transfected with expression vectors containing FOXP2 isoforms I and III and FOXP1 and FOXP4 transcripts were run on an SDS-PAGE gel, were transferred to PVDF membrane, and were hybridized with polyclonal anti-FOXP2 antibody (lanes 1–4). FOXP1 and FOXP4 were also hybridized with protein-specific antibodies (lanes 5 and 6) as a positive control for protein expression. B, FOXP2 isoform I protein immunoprecipitated from lung, BG, and IFC regions run on an SDS-PAGE gel, transferred to PVDF membrane, and hybridized with polyclonal anti-FOXP2 antibody (Ab).
The American Journal of Human Genetics  , DOI: ( /522237)
Copyright © 2007 The American Society of Human Genetics Terms and Conditions

3. Figure 2
Transcriptional targets of FOXP2 identified by ChIP-chip assay
The American Journal of Human Genetics  , DOI: ( /522237)
Copyright © 2007 The American Society of Human Genetics Terms and Conditions

4. Figure 3
Distribution of FOXP2 targets identified among tissue regions. Shown are overlapping and tissue-specific targets of the 175 BG, 144 IFC, and 192 lung target genes among the three experiments. The P values based on the hypergeometric distribution show highly statistically significant overlap between the tissues.
The American Journal of Human Genetics  , DOI: ( /522237)
Copyright © 2007 The American Society of Human Genetics Terms and Conditions

5. Figure 4
Confirmation of identified FOXP2 targets by ChIP-PCR from SH-SY5Y cells overexpressing FLAG-tagged FOXP2 (A) or IFC (B). A, Promoters of three FOXP2 targets showing enrichment of FLAG-tagged FOXP2 at their promoters compared with control immunoprecipitations (IPs). No enrichment occurred at a lung-specific promoter, ANK1, or in a gene that was not a FOXP2 target, CDH1. B, Promoters of three FOXP2 targets showing enrichment of endogenous FOXP2 compared with control IPs. Also shown is an example of an IFC target, PIR51, that did not show confirmation by ChIP-PCR. No enrichment is seen when primers are used for the ANK1 promoter or CDH1 gene. No Ab = no addition of anti-FOXP2 antibody.
The American Journal of Human Genetics  , DOI: ( /522237)
Copyright © 2007 The American Society of Human Genetics Terms and Conditions

6. Figure 5
FOXP2 overexpression confirming functional regulation of targets by FOXP2. A, Cell lysate from SH-SY5Y cells transfected with empty vector or FOXP2 isoform I, run on an SDS-PAGE gel and transferred to PVDF membrane. The membrane was hybridized with anti-FLAG antibody and anti-GAPDH antibody. B, Nineteen genes tested by qRT-PCR. The average of seven replicates and SEMs are indicated. Genes with a down-regulation in expression are shown in red, whereas those with a positive change in gene expression are shown in blue. Genes with significant difference in expression between control and FOXP2-overexpressing cells are indicated with an asterisk (*) (P<.05, by Student’s t test).
The American Journal of Human Genetics  , DOI: ( /522237)
Copyright © 2007 The American Society of Human Genetics Terms and Conditions

7. Figure 6
GO categories of in vivo targets, revealing tissue specificity of target pathways. FOXP2 target gene lists from either the CNS (BG and IFC) or lung were analyzed for significantly enriched GO categories by use of DAVID Bioinformatics Resources. Categories were considered significantly enriched if at least three genes were in one category with a P value ≤.05. Significant CNS targets with a known molecular function fall into six categories (A), whereas lung targets can be classified into three categories (B), none of which overlap with the CNS results. Significant CNS targets with a known biological function are grouped into 11 categories (C). There were no significant biological function categories for lung-specific targets.
The American Journal of Human Genetics  , DOI: ( /522237)
Copyright © 2007 The American Society of Human Genetics Terms and Conditions