Fig. 1: Mechanism of action of OTX-2002 in HCC cell lines.

A Bicistronic OTX-2002 mRNA encoding two bifunctional fusion proteins, MYC Epigenomic Controller-Element 1 (MYC-EC-E1) and Element 2 (MYC-EC-E2), target regulatory elements in the MYC Insulated Genomic Domain (IGD). Red boxes depict CCCTC-Binding factor (CTCF) binding sites of the looping IGD. Further details are available in the main text and ref. ¹⁷. B In vitro evaluation (at 48 h) of MYC-EC-E1 and MYC-EC-E2 alone or in combination compared to GFP mRNA in Hep 3B cells. Results are from one independent experiment (n = 3 technical replicates) and were repeated once. C Dose response of MYC mRNA expression and cell viability at 72 h of OTX-2002-DS treatment in three HCC cell lines. Results are from one independent experiment (n = 2 technical replicates per concentration) and were repeated once. D Annexin V/PI staining (gating strategy Fig. S1B) showing percentage of apoptotic cells in HCC cell lines treated with either negative control (NC) mRNA or OTX-2002-DS for 48 h. Results are from one independent experiment (Hep 3B, n = 6 technical replicates for all groups; Hep G2 and SK-HEP-1, n = 3 technical replicates for untreated, n = 2 technical replicates for NC mRNA, n = 4 technical replicates for OTX-2002-DS) and were repeated once. E Dose response of MYC mRNA expression and cell viability at 72 h of OTX-2002 treatment in primary human hepatocytes (PHH). Results are from one independent experiment (n = 3 technical replicates) and were repeated once. F PS: Annexin V binding showing relative luminescence of apoptotic PHH after 48 h of treatment. Results are from one independent experiment (n = 6 technical replicates for untreated, n = 3 technical replicates for NC mRNA and OTX-2002-DS) and were repeated once. For all relevant panels error bars represent mean (±) standard error of the mean (SEM). Source data are provided as a Source Data file.