OAC2 is an Oct4 activator which activates expression through the Oct4 gene promoter; enhances reprogramming efficiency by increasing the rate of production of induced pluripotent stem cells (iPSCs) from embryonic fibroblasts; an analog of OAC1.

Octamer-binding transcription factor 4 (Oct4) is crucial for initiating and maintaining cellular pluripotency, especially during early differentiation stages, and cannot be substituted by any family members for this function[1]. Essential for embryonic stem cell (ESC) pluripotency and reprogramming, Oct4, along with Sox2, Klf4, and c-Myc, benefits from the structural analog OAC2, which activates Oct4 and Nanog reporters like OAC1. OAC1 and its analogs OAC2 and OAC3 significantly enhance reprogramming efficiency by up to 2.75% and accelerate iPSC colony formation by 3 to 4 days[2].

The Oct4-luc or Nanog-luc cells are treated with compound OAC1 or its structural analogs OAC2, OAC3 at 1 μM concentration or at indicated concentrations. Other compounds used include 2 μM BIO, 2 μM BIX, 2 μM 5'-azacytidine, 25 μg/mL Vitamin C, 10 nM Am580, 5 μM tranylcypromine, and 0.5 mM valporic acid. Luciferase reporter assays are performed 24 h after compound treatment or at indicated time points. For Topflash reporter assays, 0.2 μg β-catenin–responsive Topflash reporter gene plasmid is introduced into CV1 cells using trasfection. Compounds are added 6 h after transfection. Luciferase activity is measured 48 h after compound treatment using the Glo Luciferase Assay System[2].