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. 2018 Aug 28;7:e37078. doi: 10.7554/eLife.37078

Figure 3. SE ensures efficient pol II association to intronless genes.

(A,B) Analysis of pol II occupancy at SE target loci by pol II-ChIP qPCR in WT, se-1 and se-3 in gene bodies of five different genes (A) or at various genomic regions at At3g19030 (B). Regions tested are depicted in panel D. A general pol II CTD antibody and mouse IgGs (as negative control) were used for immunoprecipitation. Additional gene loci were tested and are shown in Figure 3—figure supplement 1. Error bars indicate the range of two independent biological experiments. (C,D) Analysis of Ser5P and Ser2P pol II levels at SE target loci by ChIP qPCR in WT and se-3. General pol II CTD, pol II CTD Ser2P and pol II CTD Ser5P specific antibodies and mouse IgGs (as negative control) were used for immunoprecipitation. Additional gene loci were tested and are shown in Figure 3—figure supplement 3. Error bars indicate the range of two independent biological experiments.

Figure 3.

Figure 3—figure supplement 1. SE is important pol II association to its target genes, but does not affect pol II levels.

Figure 3—figure supplement 1.

(A–D) Analysis of pol II occupancy at SE target loci by pol II-ChIP qPCR in WT, se-1 and se-3. A general pol II antibody and mouse IgGs (as negative control) were used for immunoprecipitation. Error bars denote the range of two biological replicates. (E) Protein blot analysis of pol II levels in WT, se-1 and se-3 using pol II antibodies. ACTIN (ACT) served as a loading control.
Figure 3—figure supplement 2. The stability of RNAs produced by SE target genes is not affected SE.

Figure 3—figure supplement 2.

Analysis of RNA stability in WT, se-1 and se-3 mutants. Plants grown in liquid culture were treated with 200 µg/ml cordycepin for 0.5 hr, 1 hr and 2 hr to block transcription. Mock treatment served as a control. Quantification of indicated RNA transcript level was performed by qPCR. Error bars denote the standard error of 3 biological replicates.
Figure 3—figure supplement 3. SE ensures efficient association of pol II CTD-Ser2P and pol II CTD-Ser5P to its target genes.

Figure 3—figure supplement 3.

(A–D) Analysis of pol II occupancy at SE target loci by pol II-ChIP qPCR in WT and se-3. General pol II CTD, pol II CTD-Ser2P and pol II CTD-Ser5P specific antibodies and mouse IgGs (as negative control) were used for immunoprecipitation. Error bars denote the range of two biological replicates.