Rho action interferes in the cotranscriptional deposition of THO subunits packaging factors. (A) PMA1 ChIP of cells carrying myc-tagged THO subunits (Tho2, Hpr1, Mft1, and Thp2). The cells either did (+Rho) or did not (−Rho) express Rho. The panel shows the averages of three experiments; error bars in the graph represent SD, while asterisks denote the calculated P-value (***) P < 0.001, (**) P < 0.01, (*) P < 0.05. The top right inset illustrates the hybridization position of the oligonucleotides used for the qPCR (thick black line; coordinates on chromosome 7: 481431–481656). (B) PMA1 ChIP results for myc-tagged Tho2 strain harboring an empty vector (−Rho) or Rho expression vector (+Rho) with and without RNase A treatment. The average result for samples untreated by RNase is set as a reference at 100% for each gene locus (middle and 3′end). Percentage after RNase treatment was calculated relative to the untreated samples. The top right inset illustrates the hybridization position of the oligonucleotides used for the qPCR (middle, thick black line—coordinates on chromosome 7: 481431–481656; thick gray line—coordinates on chromosome 7: 480135–480416). The average of at least two independent experiments is shown. Error bars in the graph represent SD, while asterisks denote the calculated P-value (***) P < 0.001, (**) P < 0.01, (*) P < 0.05. (C) PMA1 ChIP and RNA immunoprecipitation experiments were performed with a myc-tagged Tho2 strain harboring an empty vector (−Rho) or a Rho expression vector (+Rho). The panel shows the averages of three experiments; error bars in the graph represent SD, while asterisks denote the calculated P-value (**) P < 0.01. The top right inset illustrates the hybridization position of the oligonucleotides used for the qPCR (thick black line; coordinates on chromosome 7: 481431–481656).
