| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | At 100% confluence, culture medium was added the adipogenic cocktail including insulin (1 μg/ml), dexamethasone (0.1 μg/ml), and IBMX (27.8 μg/ml) for 3 days, then switched to insulin (1 μg/ml) for 3 days. Plasmids pcDNA3-HA + WT GADD45A (Addgene plasmid #24929) and ING1 (Myc-DDK-tagged) (ORIGENE, #MR225175), CRISPR plasmids (synthesized by GenScript) targeting Rara, Ppard, and Ing1, and Gadd45a siRNA (BioNEER, #1360555) were transfected using a Neon Transfection System for Electroporation (ThermoFisher Scientific). | Get A Quote |
| Plasmid DNA Preparation | Get A Quote |
Retinoic acid (RA)， a bioactive metabolite of vitamin A， is a critical mediator of cell differentiation. RA blocks adipogenesis， but mechanisms remain to be established. ZFP423 is a key transcription factor maintaining white adipose identity. We found that RA inhibits Zfp423 expression and adipogenesis via blocking DNA demethylation in the promoter of Zfp423， a process mediated by growth arrest and DNA-damage-inducible protein alpha (GADD45A). RA induces the partnering between retinoic acid receptor (RAR) and tumor suppressor inhibitor of growth protein 1 (ING1)， which prevents the formation of GADD45A and ING1 complex necessary for locus-specific Zfp423 DNA demethylation. In vivo， vitamin A supplem... More
