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. 2017 Jan 4;13(1):e1006096. doi: 10.1371/journal.ppat.1006096

Fig 9. Chromatin immunoprecipitation assay showing binding of AtrR protein to the promoter regions of cyp51A and cdr1B.

Fig 9

(A) Chromatin immunoprecipitation (ChIP) was done using a mouse monoclonal antibody directed against the HA epitope. Crosslinked chromatin was prepared from a WT strain or an isogenic strain expressing a 3x HA-tagged form of atrR (shown as atrR-3× HA). Immunoprecipitated DNA was examined by semiquantitative RT PCR analysis using primers that amplified the cyp51A and cdr1B promoters. The relative enrichment of the act1 promoter was also assessed as a control for a promoter not responsive to AtrR. (B) Quantitative analysis for enrichment of the cyp51A and cdr1B promoters by real-time RT PCR method. Chromatin immunoprecipitated from the atrR-3× HA strains exhibited a 20-fold enrichment of cyp51A promoter DNA along with a ~33-fold enrichment of cdr1B promoter sequence. No detectable enrichment of act1 was found. Data are presented as the mean and standard error of two separate ChIP experiments. % input represent signals obtained from the ChIP that are divided by signals obtained from the input sample.