ASO Transfections
CECs were transfected with ASOs complexed with DarmaFECT 4 transfection reagent (Dharmacon), in accordance with the manufacturer’s instructions. The following ASOs were used; Control ASO: 5′-mG∗mG∗mU∗mG∗mG∗mA∗mU∗mC∗mA∗mC∗mG∗mA∗mG∗mU∗mU∗mC∗mA-3′, (CAG)7 therapeutic oligo: 5′-mC∗mA∗mG∗mC∗mA∗mG∗mC∗mA∗mG∗mC∗mA∗mG∗mC∗mA∗mG∗mC∗mA∗mG∗mC∗mA∗mG-3′. m denotes 2′-O-Methyl ribonucleotide, ∗ denotes phosphthioate linkages.
A final oligo concentration of 200 nM was selected for all experiments based on optimization data presented in Figure S1. All ASO treatment experiments were performed 24 hr post transfection.