a: THL does not inhibit eukaryotic RNA polymerases in vitro. Purified yeast RNA polymerases I, II and III were pre-incubated with DMSO (Ctr.), THL and α-amanitin, when indicated. Radiolabelled transcripts were synthesized from tailed DNA templates (see Methods) and analyzed by gel electrophoresis and autoradiography.
b: THL is not a Pol II inhibitor in vivo. Heat map analysis of transcriptional profiles (RNAseq) after treatment of N.crassa with 50 μM THL for 0 min, 40 min and 120 min, respectively. Yellow: high and blue: low relative expression.
c: Examples of transcription profiles (Wig files and Browser view) of N.crassa genes up- and downregulated by treatment with 50 μM THL for the indicated time periods. The number in top right corner of each panel specifies the scale (sequence reads). Scale on the top of the panels indicates 500 bp increments.
d: Pol II S5-P occupancy (ChIP-PCR) in the presence of 50 μM THL is significantly decreased at the promoters of vvd and actin but is increased at hsp98 and hsp60. Error bars indicate ±s.d. from three independent experiments measured in triplicates.