MEFs were transduced with lentiviral vectors, one encoding the transcription factors PDX1, NEUROD1 and MAFA in a polycistronic transcript, and the other mouse GLIS3-ΔN155 (Beak et al., 2008; Table S3). Cells were harvested 48 hours post transduction and subjected to quantitative PCR analysis. HEK293T, MCF7 and SW480 cells were transfected using Lipofectamine 2000, following manufacturer’s instructions. We note that the omission of MAFA or substitution of NEUROD1 with NEUROG3 in such experiments resulted in similar INS induction levels. For microarray hybridization experiments described in Figure S5, harvested RNA was hybridized to Gene ST 1.0 Affymetrix arrays and the data was analyzed on Affymetrix TAC (v1.0.24) software as described (Pasquali et al., 2014).

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.