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. 2014 Jul 8;9(7):e101969. doi: 10.1371/journal.pone.0101969

Figure 3. Pyrvinium inhibits WNT signaling in a CK1α dependent manner.

Figure 3

A. The expression of the WNT target gene: AXIN2 was determined from NIH 3T3 cells expressing control or CK1α-specific shRNAs. Cells were treated with control or WNT3a conditioned media with or without 100 nM Pyrvinium. AXIN2 expression was quantified using real-time RT-PCR. Error bars indicate ± S.E.M of three experiments. *p<0.05. B. Pyrvinium suppressed WNT signaling biomarkers in a CK1α dependent manner. Upper left, CK1α expression was assessed by real-time RT-PCR in control shRNA or CK1α shRNA expressing HCT116 cells. The three other panels show control shRNA or CK1α shRNA infected HCT116 cells treated with or without 100 nM Pyrvinium for 24 hours and AXIN2, DKK1 and LGR5 expression. Error bars indicate ± S.E.M of three experiments. *p<0.05. C. Overexpression of CK1α inhibits WNT reporter activity and decreases steady state levels of PYGOPUS2 in CRC cells. SW480 and HCT116 WTKO cells harboring the TOPflash reporter were transfected with a control plasmid or one expressing HA-CK1α. Immunoblots show decreased PYGOPUS2 levels for SW480 and HCT116 WTKO cells. Error bars indicate ± S.E.M of three experiments. *p<0.05. D. HCT116 stably expressing the indicated shRNA were treated with or without 100 nM Pyrvinium followed by determination of their colony forming capacity. Error bars indicate ± S.E.M of three experiments. *p<0.05.