Figure 4.
Functional consequences of syntenin knockdown. (A) Western blot of keratinocytes transduced with empty vector (ev) or syntenin RNAi (Rnai1) probed with anti syntenin (α-syn) or, as a loading control, anti tubulin (α-tub). (B) Hes1 luciferase reporter activity in J2-3T3 cells transduced with empty vector (ev) or the Delta constructs shown, in the presence or absence of syntenin RNAi. Fold induction relative to Renilla control is shown. Mean of 3 experiments ± SEM. Values significantly different from ev control or Fl are marked with asterisks (student’s t-test: P<0.001*** or P<0.05**). (C) Clonal growth of keratinocytes transduced with DeltaFl or Delta DS and control (Ctrl) or syntenin Rnai1. (D-G) GFP labelled clones formed by human keratinocytes expressing DeltaFl alone (Fl) (D) or in combination with syntenin Rnai1(E), or formed by mouse keratinocytes with endogenous Delta1 levels (Dll1flox/flox)(F) or lacking Delta1 (K5Cre × Dll1flox/flox)(G). Clones were photographed 5 days after seeding within a confluent sheet of unlabelled WT keratinocytes (human or mouse). Scale bar: 100 μm. (H) Percentage of cohesive colonies formed by GFP labelled keratinocytes. Data are mean of 3 experiments ± SD (student’s t-test: P < 0.001***).