Fig. 4: Identification of the lead sequence binding to SCAF8 and blocking its interaction with RNAP2.

a Docking scores of 5-nt sequences to SCAF8 generated by the Glide pipeline. The top 20 sequences are enlarged in the inserted panel. Source data are provided as a Source Data file. b Assessments of the inhibitory efficacy of the top 20 sequences on the interaction between SCAF8 and RNAP2 via competitive fluorescent polarization assay. c Evaluation of the direct interactions between the top 20 sequences and SCAF8 using SPR spectroscopy. RU, response units. d Selection of DNA sequences exhibiting both binding affinity and inhibitory efficacy. e Standard kinetics SPR assay demonstrating the binding of SCAF4_LS (D2_5-nt) to immobilized SCAF. This experiment is performed with three biological replicates. Average affinity value is 76.40 ± 7.33 nM. Affinities of each replicate are indicated in the bracket (biological replicates n = 3, mean ± SD). Source data are provided as a Source Data file. f Dose-response curves of SCAF4_LS (D2_5-nt) depicting the inhibition of RNAP2 interaction with SCAF8. The IC₅₀ is determined from three biological replicates, each containing three technical replicates. The averaged IC₅₀ value is calculated from all experiments indicated in brackets (biological replicates n = 3, mean ± SD). Source data are provided as a Source Data file. g–k SPR results illustrating the binding of base group-deleted SCAF4_LS (D2_5-nt) sequences to SCAF8. Average affinities are generated from three independent experiments, with affinities of each experiment indicated in the bracket. Source data are provided as a Source Data file.