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1.
Figure 2.

Figure 2. From: Genome-wide identification of in vivo protein-DNA binding sites from ChIP-Seq data.

Distribution of binding sites across the genome. RefSeq genes were used as reference. The 5 kb region upstream of transcription start site was defined as the promoter.

Raja Jothi, et al. Nucleic Acids Res. 2008 September;36(16):5221-5231.
2.
Figure 3.

Figure 3. From: Genome-wide identification of in vivo protein-DNA binding sites from ChIP-Seq data.

Accuracy of inferred binding sites. Consensus binding motifs for CTCF, NRSF and STAT1 binding sites are shown in the left panel. The right panel shows the percentage of identified binding sites containing the consensus binding motif(s).

Raja Jothi, et al. Nucleic Acids Res. 2008 September;36(16):5221-5231.
3.
Figure 5.

Figure 5. From: Genome-wide identification of in vivo protein-DNA binding sites from ChIP-Seq data.

Tag density at a binding site is an indicator of binding affinity. (A) Distribution of the number of tags within the 200-bp region centered on binding sites exhibits a power-law behavior. (B) Motif analysis on four mutually exclusive sets of 500 NRSF binding sites each reveals a positive correlation between the information content of first 6 nt of the NRSF consensus motif and the average tag density. See Supplementary Figure 2 for an analysis of CTCF binding sites.

Raja Jothi, et al. Nucleic Acids Res. 2008 September;36(16):5221-5231.
4.
Figure 1.

Figure 1. From: Genome-wide identification of in vivo protein-DNA binding sites from ChIP-Seq data.

Schematic overview of SISSRs algorithm. (A) Sequenced short reads (typically ∼25–50 bp) from ChIP-Seq experiments are first mapped onto the reference genome. The mapped reads are then used to estimate statistical parameters, which include the estimation of the average length F of sequenced DNA fragments. (B) The entire reference genome along with mapped reads is scanned using overlapping windows of size w base pairs (overlapping not shown in the figure for clarity), and the net tag count (ci) for every window i is calculated. Every transition point (t) is a candidate binding site, and needs to satisfy a set of estimated as well as user-set thresholds in order to be classified as a true binding site.

Raja Jothi, et al. Nucleic Acids Res. 2008 September;36(16):5221-5231.
5.
Figure 6.

Figure 6. From: Genome-wide identification of in vivo protein-DNA binding sites from ChIP-Seq data.

Overcoming ChIP-Seq limitations. (A) Nonspecific reads due to antibody non-specificity and/or sequencing errors could be controlled for by adjusting the window size w. (B) SISSRs provides an option to identify binding sites with tags mapped to only the sense or antisense strand, which otherwise may not have been identified (site A). Steps are taken to make sure that tags corresponding to site B are not accounted while identifying site A: the right most sense tag i for site A should be at least a fragment length F away from the nearest sense tag (j) and at least 2F away from the nearest antisense tag k. (C) False positives due to indirect DNA binding via protein–protein interaction is a cause for concern, and are difficult to avoid as it is very difficult to determine whether the binding to the DNA is direct or indirect.

Raja Jothi, et al. Nucleic Acids Res. 2008 September;36(16):5221-5231.
6.
Figure 4.

Figure 4. From: Genome-wide identification of in vivo protein-DNA binding sites from ChIP-Seq data.

Resolution of inferred binding sites. (A) Distribution of the distance between SISSRs-inferred binding site and the middle of the nearest canonical motif occurrence. The red curve is for all binding sites, and the blue curve is for high-scoring binding sites. About 75% of all CTCF, NRSF and STAT1 sites were within 18, 27 and 51 bp of the nearest consensus motif, respectively (90% within 31, 52 and 73 bp, respectively). The resolution is much higher for high-scoring binding sites (90% within 20, 19 and 45 bp, respectively). (B) Distribution of the number of canonical motifs within the 200-bp region centered on identified binding sites. The coloring scheme is the same as that used in A. (C) A 200-bp region centered on a CTCF binding site (highlighted in black) with high tag density. Nine instances of core CTCF motif (10−8 < P < 0.0025) are highlighted in yellow.

Raja Jothi, et al. Nucleic Acids Res. 2008 September;36(16):5221-5231.

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