Figure 8

Inhibition of the nuclear translocation of phospho-β-catenin (Ser675) by RS-PP-050. (A) Immunoblot representing the decrease in protein expression of phospho-β-catenin (ser675) in HT-29 cells after treatment with RS-PP-050 for 24 h. β-actin serves as a loading control. (B) Bar graph represents the normalized band intensities of phospho-β-catenin (ser675) to total β-catenin. Data are means ± S.E.M compared with the vehicle control (n = 4) (**P < 0.01). (C) Immunoblot representing the inhibition in protein expression of phospho-β-catenin (ser675) in nuclear extracts of HT-29 cells after treatment with RS-PP-050 (10 μM) for 24 h. α-Tubulin, and lamin A/C serve as a cytosolic and a nuclear marker, respectively. (D) Bar graphs represent the normalized band intensities of phospho-β-catenin (ser675) to α-Tubulin, and lamin A/C. Data are means ± S.E.M compared with the vehicle control (n = 3) (**P < 0.01). For the cropped blots in Fig. 8A,C, protein samples were run under same conditional treatments and processed in parallel. Full-length blots are presented in Supplementary Fig. S3. (E) Immunofluorescent image demonstrating the reduction of nuclear accumulation of phospho-β-catenin (ser675) (green) in HT-29 cells after treatment with RS-PP-050 (5 μM) for 12 h (Lower panel). DAPI was used as a nuclei marker (blue). The slides were visualized with a confocal laser microscopy. Scale bars = 20 μM.