Fig. 3: Rio1 downregulates RNAPII recruitment and activity at (peri)centromeres.

a CenRNA levels measured by RT-qPCR analysis in wild-type yeast (RIO1), in the RIO1-AID strain treated with 500 µM auxin or a mock, in a cbf1∆ mutant, and in a cbf1∆ RIO1-AID strain treated with 500 µM auxin or a mock. Transcript concentrations were normalised to those of ACT1 and then referenced to the corresponding cenRNA levels measured in the wild-type strain (value = 1). Singular data (white circles) deriving from five biological replicates (n = 5) are shown combined as mean ± SEM. P-values were calculated with the unpaired, two-tailed Student’s t test. b RNA polymerase II levels at CEN5-periCEN5 and CEN8-periCEN8 as determined by anti-RNAPII ChIP-qPCR analysis in early S-phase RIO1-AID cells (following α-factor arrest-and-release) in the presence of 500 µM auxin or a mock. The data were normalised to those measured in the mock-treated cells (four replicate experiments (n = 4), shown as mean ± SEM). P-values were calculated with the unpaired, two-tailed Student’s t test. c RNA polymerase II levels at CEN5-periCEN5 and CEN8-periCEN8, measured by anti-RNAPII ChIP-qPCR analysis in G1, S-phase, and metaphase RIO1-AID cells, following α-factor arrest-and-release in the presence of 500 µM auxin or a mock. The data were normalised to those measured in G1 (α-factor arrested cells). The results derived from five replicate experiments (n = 5) and are shown as mean ± SEM. P-values were calculated with the unpaired, two-tailed Student’s t test. d Cen5 and pericen5 RNA levels measured by RT-qPCR analysis in RIO1 and RIO1-AID cells treated with a mock, 3 µM thiolutin and/or 500 µM auxin. The cells were released from G1 and analysed in early S-phase (five experimental replicates per condition, n = 5). Transcript levels were normalised to those measured in the untreated wild-type strain at T = 0 min. The singular data (white circles) are shown combined as mean ± SEM. P-values were calculated with the unpaired, two-tailed Student’s t test.