Fig. 7.

Partial inhibition of E-cadherin expression with siRNA interferes with SMG branching morphogenesis and normal duct development. E12.5 SMGs were transfected with either E-cadherin siRNA or with a non-silencing control and grown in culture for 22– 48 hr. A: Total RNAs, isolated from SMGs treated with either E-cadherin siRNA or non-silencing controls, were analyzed by real-time PCR and normalized to 29S. Values are expressed as a relative fold change compared to control. E-cadherin mRNA levels were reduced 30% following treatment with E-cadherin siRNA compared to controls. P values are unpaired t-tests compared to 29S controls, **P < 0.05. B: Phase microscopy of SMGs transfected with non-silencing (NS) and E-cadherin siRNAs (S) (a,b). The number of buds in E-cadherin siRNA-treated glands was reduced by 55% compared to non-silencing controls (c) (ANOVA, **P < 0.05). C: Confocal imaging of F-actin and nuclei of E12 SMGs transfected with either non-silencing control (a– c) or E-cadherin siRNA (d–f) for 22 hr revealed dilated ducts in the E-cadherin siRNA treated glands (d). Width of ductal lumens is denoted by brackets in a and d). Scale bar = 10 μm.