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. 2011 Jun 20;6(6):e21172. doi: 10.1371/journal.pone.0021172

Figure 3. Dia2 is bound to chromatin during transcription.

Figure 3

3A) Dia2 binds to the HTA1/HTB1 promoter. Chromatin immunoprecipitation was performed using haemagglutinin epitope-tagged Dia2 (Dia2-HA) and untagged control strains. Precipitated DNA was analysed by quantitative PCR using primers spanning the HTA1/HTB1 promoter (A to E) as depicted (top). qPCR analysis of Dia2 recruitment to the HTA1/HTB1 locus in the absence (Asyn.) or presence of hydroxyurea (HU) are plotted (bottom). Each data point is normalised to its corresponding input. Signals obtained from an untagged control sample are plotted on the right for comparison. ‘ORF’ denotes a primer located within the HTA1 open reading frame. ARS denotes primers amplifying an origin of replication (ARS428) close to HTA1/HTB1. 3B) Dia2 is recruited to the GAL1 promoter and ORF during gene induction Chromatin immunoprecipitation of HA-tagged Dia2 was carried out in wild-type cells grown either in raffinose or 20 minutes after galactose induction. Positions of primers (A and B) for GAL1 used to amplify the precipitated DNA are depicted in the top panel. Dia2 binds to GAL1 only during gene induction (in galactose but not in raffinose). In the presence of raffinose, the level of Dia2 recruitment is insignificant (comparable to background levels in the untagged control). Each data point is normalised to its corresponding input. Signal obtained from an untagged control strain are depicted on the right for comparison. ‘Con’ denotes a transcriptionally silent subtelomeric region on chromosome II.