Figure 4From: A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in DrosophilaCell 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.Back to article page