Figure 5: Verification of putative VRN1-HA-binding sites by ChIP-PCR.
From: Direct links between the vernalization response and other key traits of cereal crops
(a) ChIP-PCR was used to verify 10 potential VRN1-HA-binding sites using VRN1-HA line 6 and a sibling null control line. Peak numbers correspond to those in Supplementary Data 2. (b) ChIP-PCR data from multiple sites of the promoter of ODDSOC2 in lines carrying the VRN1-HA construct versus sibling null controls. Assay was performed in cv. Golden Promise and in a vernalization-responsive version of the same cultivar, ‘Winter Golden Promise’. Target region is shown in grey. Thick line indicates first exon, medium line indicates transcribed regions and thin lines indicate 5′ upstream sequence. The location of potential MADS box transcription factor-binding sites is indicated in red (CArG box). (c) The binding of VRN1-HA to VRN2a and VRN2b (duplicate genes at the VRN2 locus of barley) assayed by ChIP-PCR. VRN1-HA binding was assayed in the Winter Golden Promise background. VRN1 binding indicates relative enrichment, immunoprecipitated DNA versus input chromatin, for three biological repeats. Error bars show s.e. Stars indicate statistical significance by Student’s t-test: *P<0.05, **P<0.01, ***P<0.001.
