Fig. 2

shRNA oligo design and loop comparison. a Format for shRNA oligo design. Upper strand is sense oligo, lower strand is anti-sense oligo. b Diagram of predicted shRNA loop structure with a basic SpeI sequence (6 nt: ACUAGU) or including a single stem mismatch (7 nt: UACUAGU). Colors correlate to calculated likelihood of the depicted pairing. See methods for details on prediction tool. c Immunoblot showing two different pools of iPrEC cells with shRNA against p38δ, with the only difference being a single mismatch in the loop sequence of the shRNA. Cells were treated −/+ Dox for 72 h. TetR was probed on a separate gel. p38α and Tubulin serve as loading controls. d Same experiment as c using a different pair of shRNAs targeting Creb1. Cells were treated −/+ Dox for 5 days