Scientists use AI to identify likely drug targets in search for Alzheimer's cure CommsBio
, with minor modifications. Briefly, iPSCs were plated in mTeSR1 media at a density of 95 K cells/cmon Matrigel-coated plates for viral transduction. Media was changed from StemFlex to mTeSR1 as we found better transduction viability with mTeSR1. Viral plasmids were obtained from Addgene . FUdeltaGW-rtTA was a gift from Konrad Hochedlinger , and Tet-O-FUW-EGFP and pTet-O-Ngn2-puro were gifts from Marius Wernig.
Briefly, conditioned media from transduced cells were incubated in pre-blocked wells along with detection antibody solution. Plates were read using an MSD SECTOR Imager 2400 and resulting peptide concentrations were normalized to total protein in the cell lysate per well measured using the Pierce BCA Protein Assay Kit . Data for each shRNA knockdown were additionally normalized to the average of control conditions for each parameter measured.