Dibenzazepine dbz

Dibenzazepine (DBZ) was purchased from Cayman. In vivo, mice received intraperitoneal injections every other day consisting of 100μl of a solution of GSI (0.1mg) dissolved in DMSO and olive oil or vehicle alone (DMSO with olive oil). Mice were randomized into treatment groups. DMSO and GSI-treated animals were co-housed. In vitro, GSI was dissolved in DMSO and was added to the culture media at a final concentration of 0.01μM. Control cells were incubated with an equal concentration of DMSO. Cells were treated with GSI or DMSO for 30 minutes prior to stimulation.

To detect the drug effects on ACD, BAM‐transduced fibroblasts were treated with the PKCζ pseudo‐substrate inhibitor (Cayman chemical, 799764‐07‐1) at the indicated concentrations for 24 h prior to the addition of Dox. The inhibitor was then administered in a MEF medium containing Dox for 24 h followed by the neuronal induction medium for the remainder of the experiment. Parallel conditions with DMSO served as a control. ACD was detected via EdU staining based on a 3‐h EdU labeling, and ACD percentage was determined as the percentage of asymmetrically dividing cells after Dox treatment for 24 h (unless otherwise indicated) relative to the number of cells seeded. Similar experiments were performed using the Notch inhibitor, dibenzazepine (DBZ; Cayman chemical, 14 627).
To determine the effect of cell cycle inhibition on iN reprogramming, CDK4 inhibitor (1µm; Cayman chemical, 546102‐60‐7) was used to inhibit the cell cycle in the G0/G1 phase. BAM‐transduced fibroblasts were treated with the small molecule inhibitors (i.e., CDK4 inhibitor and PKCζ pseudo‐substrate inhibitor) either on day ‐1 (i.e., in MEF medium and 12 h before the addition of Dox) or day 1 (i.e., in N2B27 medium and 24 h after the addition of Dox) and then throughout the first week of reprogramming. On day 5, samples were fixed, stained, and the reprogramming efficiency was determined.