Figure 6. CRIP1 activates the Wnt/β‐catenin pathway by decreasing β‐catenin acetylation.

• A
  ELISA analysis of acetylcarnitine levels in CRIP1‐knockdown cells treated with mildronate (5 mM) for 24 h.
• B
  GSEA based on CRIP1 expression in the TCGA‐LIHC database.
• C, D
  Representative images of tumor sphere formation and quantification of spheres (C) and flow cytometric analysis of the proportion of CD133⁺ cells (D) in CRIP1‐overexpressing Huh7 cells with or without iCRT3 (50 μM) for 24 h. Scale bar: 50 μm.
• E
  Western blot analysis of HCC cancer stem cell markers in CRIP1‐overexpressing Huh7 cells with or without iCRT3 (50 μM) for 24 h.
• F
  Co‐immunoprecipitation and immunoblot analyses of β‐catenin acetylation in CRIP1 knockdown MHCC‐97H cells treated with mildronate (5 mM) for 24 h.
• G
  Western blot analysis of cytoplasmic and nuclear fractions of β‐catenin in CRIP1 knockdown MHCC‐97H cells treated with mildronate (5 mM) for 24 h.
• H
  Confocal microscopy images of β‐catenin in CRIP1 knockdown MHCC‐97H cells treated with mildronate (5 mM) for 24 h. Scale bar: 10 μm.
• I
  Co‐immunoprecipitation and immunoblot analyses of β‐catenin ubiquitination in CRIP1 knockdown MHCC‐97H cells treated with mildronate (5 mM) and EX527 (20 μM) for 24 h.

Data information: Data represent the mean ± SD of at least three independent experiments. **P < 0.01 and ***P < 0.001. Differences were tested using an unpaired two‐tailed Student's t‐test (A, C and D).