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. 2022 Jul 1;41(15):e110218. doi: 10.15252/embj.2021110218

Figure 5. BBOX1 ubiquitination at K240 mediated by STUB1 promotes cancer stem‐like properties induced by CRIP1.

Figure 5

  • A
    BBOX1 ubiquitination modification at Lys240 by MS/MS.
  • B
    Co‐immunoprecipitation and immunoblot analyses of BBOX1 ubiquitination in STUB1‐overexpressing Huh7 and Hep3B cells transfected with BBOX1‐WT or BBOX1 K240R mutant.
  • C
    Western blot analysis of BBOX1‐WT or K240R mutant protein expression in Huh7 and Hep3B cells transfected with increasing amounts of STUB1.
  • D‐G
    Carnitine levels (D), tumor sphere formation ability and quantification of spheres (E), flow cytometric analysis of the proportion of CD133+ cells (F), and protein levels of HCC cancer stem cell markers (G) in STUB1‐overexpressing Huh7 and Hep3B cells transfected with BBOX1‐WT or BBOX1 K240R mutant. Scale bar: 50 μm.

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 (D–F).