Confirmation of the effects of CS-E on Wnt3a target gene expression by qRT-PCR. NIH3T3 cells were treated with L-CM (L) or Wnt3a-CM (W3a) in the presence or absence of CS-E at 100 μg/ml for 36 h followed by RNA preparation and qRT-PCR. A, evaluation of effects of CS-E treatment on expression of the positively regulated target genes Calr, Arhgap18, Prickle1, Tnfsf9, and Mrgprf in the presence of L-CM or Wnt3a-CM. All five target genes were confirmed to be induced by treatment with Wnt3a-CM. Concomitant treatment with CS-E resulted in a significant decrease in expression of Calr, Arhgap18, Prickle1, Tnfsf9, and Mrgprf (*, p < 0.05). B, evaluation of effects of CS-E treatment on expression of the negatively regulated target genes Cyp2f2, Slpi, Arrdc4, Lcn2, and Sox9 in the presence of L-CM or Wnt3a-CM. All five target genes were confirmed to be repressed by treatment with Wnt3a-CM, and concomitant treatment with CS-E did not result in any significant change in Wnt3a-mediated repression in any of the target genes.