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. 2016 Jun 15;7(29):46187–46202. doi: 10.18632/oncotarget.10070

Figure 3. Effects of Wnt3a and Wnt5a stimulation and FZD2 blockade on cell proliferation, migration and Rac1 activity in NB cells.

Figure 3

A. Relative density of cancer cells up to 72 h following stimulation with 100 ng/ml recombinant Wnt3a, Wnt5a or pretreatment with FZD2 siRNA (FZD2si) or nonspecific scrambled siRNA (scrCo) was measured using the WST-1 cell proliferation assay. FZD2 siRNA suppressed NB cell proliferation. Graphs represent the mean of 3 independent experiments ± SD. Asterisks (*) indicate P < 0.04 vs. untreated controls (Co, scrCo). B. Representative images of migrated SK-N-AS and SK-N-DZ NB cells from an in vitro migration assay are shown (200x magnification). C. Quantification of cell migration. NB cells were stimulated with 100 ng/ml recombinant Wnt3a and Wnt5a with or without pretreatment with FZD2 siRNA or scrambled siRNA (scrCo) for 24 h and migrated cancer cells quantified subsequently by in vitro migration assays. FZD2 blockade suppressed Wnt-induced migration of NB cells. Graphs represent the mean of 3 independent experiments ± SD. Asterisks (*) indicate P < 0.05 vs. untreated controls (Co, scrCo) and vs. Wnt3a and Wnt5a stimulated cells, respectively. D. Rac1 activation in SK-N-AS and SK-N-DZ NB cell lines after stimulation with 100 ng/ml Wnt3a or Wnt5a protein for 30 min with or without FZD2 siRNA or scrambled siRNA (scrCo) pretreatment. FZD2 blockade suppressed Wnt-induced Rac1-activation of NB cells. Graphs represent the mean of 3 independent experiments ± SD. Asterisks (*) indicate P < 0.05 vs. untreated controls (Co, scrCo) and vs. Wnt3a and Wnt5a stimulated cells, respectively.