Figure 11

Studies on intracellular WNT-signaling in HD-LECs using small molecules in scratch-assays. (A) Scratch assay with HDLECs pre-treated with LGK974 for 3 days and subsequent application of recombinant WNT5A. Representative images after 0 h and 24 h are shown. (B) Quantification of scratch closure after 24 hours. Percentage of closure is shown. Inhibition of PORCN with 10 µM LGK974 reduces migration significantly (p < 0.0001). Application of 500 ng recombinant WNT5A increases migration (p = 0.0021). (C) Inhibition of the canonical WNT-pathway with FH535 does not affect scratch closure (p = 0.2168). Activation of the canonical WNT-pathway with GSK-3 inhibitors IM-12 or BIO inhibits migration (p = 0.0013 and p < 0.0001, respectively). (D) Inhibition of the PCP-pathway by ROCK-inhibitors Y-27632 or Fasudil does not affect scratch closure (p = 0.2904 and p = 0.3595, respectively). (E) Inhibition of the PCP-pathway by RAC-inhibitors EHT-1864 or NSC23766 reduces scratch closure (p < 0.0001 and p < 0.0001, respectively). (F) Inhibition of the PCP-pathway by JNK-inhibitors SP600125 or JNK-IN-8 reduces scratch closure (p < 0.0001 and p < 0.0001, respectively). At least 3 independent experiments; number of replications see text; t-test: all compared to control, if not stated otherwise.