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3box PSE -55 DSE -220+1 +350 U2 -800-500 +700 +150 -15-200 +215 % of input PTF 4563 Figure S1. High resolution chromatin immunoprecipitation of PTF on.

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Presentation on theme: "3box PSE -55 DSE -220+1 +350 U2 -800-500 +700 +150 -15-200 +215 % of input PTF 4563 Figure S1. High resolution chromatin immunoprecipitation of PTF on."— Presentation transcript:

1 3box PSE -55 DSE U % of input PTF 4563 Figure S1. High resolution chromatin immunoprecipitation of PTF on U2 snRNA genes. qRT-PCR of ChIP analysis of the U2 gene using anti-PTF antibodies and the primers noted on the diagram of the U2 genes shown above.

2 % of input Histone H4 Histone H2A Histone H2B RNU2 locus -actin gene Histone H4 Histone H2A Histone H2B % of input Figure S2. All four histones have the same pattern of depletion on the U2 and the -actin genes. qRT-PCR of ChIP analysis of the U2 and -actin genes using antibodies against the histones noted.

3 % of input - -amanitin + -amanitin % of input Histone H3RNA pol II RNU2 locus Figure S3. Nucleosomal depletion on U2 genes is transcription-independent. qRT-PCR of ChIP analysis of the U2 gene using antibodies against H3 and pol II, with or without treatment of the cells with -amanitin.

4 3box PSE -55 DSE CT-rich +600 U transcription % of input NELF-E Histone H3 CTCF Figure S4. Fine mapping of NELF and CTCF on the RNU2 locus. qRT-PCR of ChIP analysis of the RNU2 locus, using antibodies against H3, NELF-E and CTCF using the primers noted on the diagram of the RNU2 locus above.

5 RNU2 locus -actin gene 3box PSE -55 DSE CT-rich +600 U2 4a Ser5 relative to Pol II Ser7 relative to Pol II -800 Figure S5. Distribution of Ser5 and Ser7 phosphorylation on the RNU2 locus and the -actin gene. qRT-PCR of ChIP analysis using antibodies against phospho-Ser5 and phospho-Ser7, relative to the Pol II signal. The additional primer, 4a, is noted on the diagram of the RNU2 locus above. All other primers are as shown in Figure 1.

6 Spt16 level (% input) Pol II level (% input) Spt16 Pol II Spt16 Pol II RNU2 locus -actin gene Figure S6. FACT is associated with both snRNA and protein-coding genes. qRT-PCR of ChIP analysis of the U2 and -actin genes, using antibodies against the Spt16 subunit of FACT. The pol II level is indicated by a dotted line. Ac-H4 (related to H4) RNU2 locus -actin gene H4 level (% input) AcH4/H4 H4 Figure S7. H4 acetylation across the RNU2 locus and -actin gene. qRT-PCR of ChIP analysis of the U2 and -actin genes using antibodies against H4 and acetyl H4 (Ac-H4). The H4 level is indicated by a dotted line and the Ac-H4 level is shown relative to total H4.

7 H3-K36M 3 (related to H3) RNU2 locus -actin gene +DRB DRB - +DRB DRB - H3-K36M 3 (related to H3) Figure S8. H3-K36 trimethylation is DRB-sensitive. qRT-PCR of ChIP analysis of the U2 and -actin genes using antibodies against trimethyl H3-K36 (H3-K36M3) with or without treatment of the cells with DRB. H3-K9M 3 (related to H3) RNU2 locus -actin gene H3 level (% input) H3-K9M 3 /H3 H3 H3-K9M 3 (related to H3) Figure S9. H3-K9 trimethylation across the RNU2 locus and -actin gene. qRT-PCR of ChIP analysis of the U2 and -actin genes using antibodies against trimethyl H3-K9 (H3-K9M3). The H3 level is indicated by a dotted line and the H3-K9M3 level is shown relative to total H3.

8 RNU2 locus -actin gene GAPDH gene -actin gene / / / / / / / / / / / / / / / / / / / / / / / / / / / /+3682 Figure S10. Regions analysed by qRT-PCR after chromatin immunoprecipitation. Regions amplified by each pair of primers on the RNU2 locus, the -actin, GAPDH and -actin genes are indicated relative to the transcription start site.


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