Knockdown of MAFA in EndoC-βH1 (Ravassard et al., 2011 (link)) cells was achieved using the TARGETplus siRNA pool against human MAFA (no. L-027343-01). Briefly, 2 × 106 cells were transfected with siRNA (50 pmol) or a non-targeting control (no. D001810) using the Dharmafect no. 1 reagent (GE Dharmacon no. T-2001) following the manufacturer’s protocol. RNA was harvested 72 hr post-transfection and isolated using Trizol reagent (Life Technologies) and the DNA-Free RNA Kit (Zymo Research). The iScript cDNA synthesis kit (Bio-Rad) was used for cDNA synthesis, and qPCR reactions were performed on a LightCycler 480 II (Roche) and analyzed by the ΔΔCT method.
Knockdown of MafA or control sequences in βTC-6 cells was performed using 30 nM MAFA RNAi oligonucleotides (no. s196287; Thermo Scientific) or negative control no. 1 (Thermo Scientific) with transfection reagent (Dharmafect; Thermo Scientific). Total RNA was extracted after 72 hr transfection, and qPCR was performed as described above.
Knockdown of MAFA resulted in a 50% reduction of the MAFA transcript and protein levels (data not shown).
Knockdown of MafA or control sequences in βTC-6 cells was performed using 30 nM MAFA RNAi oligonucleotides (no. s196287; Thermo Scientific) or negative control no. 1 (Thermo Scientific) with transfection reagent (Dharmafect; Thermo Scientific). Total RNA was extracted after 72 hr transfection, and qPCR was performed as described above.
Knockdown of MAFA resulted in a 50% reduction of the MAFA transcript and protein levels (data not shown).