Fig 2: SNOND15B regulates cellular sublocalization of p53. (a-b) SNORD15B expression had no effect on the total P53 level in endometrial cancer cells. (c) Knocking down TRIM25 reversed the proliferation effect of SNORD15B. (d) Overexpression of SNORD15B in Ishikawa cells reduced P53 expression in the nucleus. (e) Knockdown of SNORD15B in HEC-1B increased P53 expression in the nucleus ****P < 0.0001.

Fig 3: TRIM25 influences the sensitivity of CRC cells to oxaliplatin.A Western blotting analysis of TRIM25 in SW48 and SW480 cells transfected with shTRIM25 or a TRIM25-expressing plasmid. GAPDH was used as the loading control. B–C A CCK8 assay was used to measure the viability of the indicated cells treated with different concentrations of OXA for 48 h. D Colony-formation ability (left panel: representative images; right panel: quantification of colony numbers) of the indicated cells treated with OXA (2 µM). E Annexin V-FITC and PI staining showing apoptosis in the indicated CRC cells treated with OXA (30 µM) for 48 h. Left panel: representative images; right panel: Quantification of apoptotic cells. F Western blotting analysis of cleaved caspase 3 and cleaved PARP in the indicated cells treated with or without OXA (30 µM) for 48 h. Data are represented as the mean ±SD of three independent experiments. *P < 0.05, **P < 0.01, **P < 0.001.

Fig 4: CypA enhances TRIM25-mediated K63-linked ubiquitination of RIG-I and facilitates recruitment of RIG-I to MAVS.(A) Immunoblot analysis of lysates in 293T/CypA- cells transfected for 24 hr with Flag-RIG-I, along with HA-K63-Ub, HA-K48-Ub, Myc-TRIM25, or Myc-CypA, followed by immunoprecipitation with anti-Flag beads. (B) Immunoblot analysis of lysates in WT and Ppia-/- BMDMs infected with SeV for 6 hr, followed by immunoprecipitation with control mouse IgG or anti-RIG-I antibodies. Lysates and immunoprecipitation extracts were probed with K63-Ub, RIG-I and CypA antibodies. (C) Immunoblot analysis of lysates in 293T/CypA- cells transfected with Flag-RIG-I, Myc-TRIM25, or Myc-CypA for 24 hr, and immunoprecipitated with anti-Flag beads. (D) Immunoblot analysis of lysates in WT and Ppia-/- BMDMs infected with SeV for 6 hr, followed by immunoprecipitation with control mouse IgG or anti-RIG-I antibodies. Lysates and immunoprecipitation extracts were probed with RIG-I, TRIM25, CypA and antibodies. (E) Immunoblot analysis of lysates in 293T/CypA- cells transfected with Myc-CypA and Flag-RIG-I, Flag-RIG-I-C or Flag-RIG-I-N for 24 hr, and immunoprecipitated with anti-Flag beads. (F) Immunoblot analysis of lysates in WT and Ppia-/- BMDMs after SeV infection or mock-infection for 6 hr, followed by mitochondrial-cytoplasm extraction. (G) Confocal microscopy of endogenous RIG-I in 293T/CypA+ and 293T/CypA- cells stained with Mito-Tracker after SeV infection or mock infection for 6 hr. Scale bars, 10 µm. (H) Immunoblot analysis of lysates in 293T/CypA+ and 293T/CypA- cells transfected with Flag-RIG-I and Myc-MAVS for 24 hr, and immunoprecipitated with anti-Flag beads. (I) Confocal microscopy of endogenous CypA in 293T/CypA+ cells stained with Mito-Tracker after SeV infection or mock-infection for 6 hr. Scale bars, 10 µm. Data are representative of at least three independent experiments.DOI: http://dx.doi.org/10.7554/eLife.24425.016

Fig 5: The working model depicting the role of TRIM25 in CRC cells.TRIM25 stabilizes EZH2 by blocking TRAF6-mediated ubiquitination for EZH2 degradation, and sustains stem cell properties to promote the resistance of CRC to OXA.