Figure 2.

DsiRNAs effectively silence targets in poorly differentiated PAE cultures. (a) Poorly differentiated cultures, 2-day post-seeding, were transfected with 250 nmol/l of DsiRNA against HPRT with the use of RNAiMAX, Transductin, or Accell siRNA at the indicated concentrations and then processed (RNA isolation, RT-qPCR) 24 hours later. Remaining mRNA levels were normalized to negative control (NC1). Mean levels (±SD) were calculated from three biological replicates (in triplicate) **P < 0.01, ***P < 0.001 (Student's t-test). (b–c) Confocal images (x–z stacks) of epithelia 2 hours after transfection with DIG-HPRT DsiRNA complexed with (b) Transductin or (c) unformulated. Blue, labeled nuclei; green, DIG-labeled oligo.