Figure 4

Cell type specificity for signaling pathways. The basal activity and fold induction by Wg or ΔNLrp6 is different in the variety of Drosophila cell lines tested. (a-d) The wg reporter can be induced by the expression of both Wg and ΔNLrp6 in clone 8 and SL2 cells (a,b) but only with Wg in Kc167 and S2R+ cells (c,d). (e,f) dsRNA-mediated knockdown of known positive (e) and negative (f) regulators variably affect the activity of the Wg reporter in different cell lines. (g,h) Effect of dsRNA-mediated knockdown of DFz2 receptor in different cell types. RNAi inhibition of DFz2 inhibits Wg pathway activity in S2R+ cells (g) but not in clone 8 cells (h). (i) Western blot to detect expression of a negative regulator, Axn in clone 8, Kc167 and S2R+ cell lines. Levels of Axn in clone 8 cells is significantly lower than in S2R+ or Kc167. Anti-α-tubulin antibody was used as a loading control (i). All luciferase reporter assays were performed in 4 replicas and error bars represent the standard error between the four data points.