General Observation of Fe³⁺/Fe²⁺ Redox Couple Close to 4 V in Partially Substituted Li₂FeP₂O₇ Pyrophosphate Solid-Solution Cathodes

Exploring the newly unveiled Li₂<i>M</i>P₂O₇ pyrophosphate cathode materials for lithium-ion batteries, the current study reports the general observation of an unusually high Fe³⁺/Fe²⁺ redox potential close to 4.0 V vs Li/Li⁺ in mixed-metal Li₂<i>M</i>_<i>x</i>Fe_1–<i>x</i>P₂O₇ (<i>M</i> = Mn, Co, Mg) phases with a monoclinic structure (space group <i>P</i>2₁/<i>c</i>). Such a high voltage Fe³⁺/Fe²⁺ operation over 3.5 V has long been believed to be possible only by the existence of much more electronegative but hygroscopic anions such as SO₄^2– or F^–. Thereby, this is the first universal confirmation of >3.5 V operation by stable, simple phosphate material. High voltage (close to 4 V) operation of the Fe³⁺/Fe²⁺ couple was stabilized by all dopants, either by larger Mn²⁺ or smaller Co²⁺ and Mg²⁺ ions, where Mg²⁺ is redox inactive, revealing that the high voltage is induced neither by reduced Fe–O bond covalency nor by contamination by the redox couple of other transition metals. The cause of higher Fe³⁺/Fe²⁺ redox potential is argued and rooted in the stabilized edge-sharing local structural arrangement and the associated larger Gibbs free energy in the charged state

Additional file 4: of Ajuba inhibits hepatocellular carcinoma cell growth via targeting of β-catenin and YAP signaling and is regulated by E3 ligase Hakai through neddylation

Figure S4. Hakai mediates Ajuba degradation via neddylation. (A) Immunoblot analysis and quantification of the half-life of Ajuba in the presence of cycloheximide (CHX, 80 μg/ml), and in the presence or absence of MLN4924 (5 μM) in BEL7402 and HepG2 cells. GAPDH was used as a loading control. (B) Ubiquitination (Ub) assay of Ajuba in 293 T cells transfected with the indicated plasmids. (C) Neddylation assay of Ajuba in 293 T cells transfected with the indicated plasmids. IB, immnoblot. IP, immunoprecipitation. WCL, Whole-cell lysates. (JPG 103 kb

Additional file 6: of Ajuba inhibits hepatocellular carcinoma cell growth via targeting of β-catenin and YAP signaling and is regulated by E3 ligase Hakai through neddylation

Figure S6. Hakai promotes BEL7402 cells invasion and growth. (A) Representative images and quantification of invasion in GFP-tagged Hakai-overexpressing BEL7402 cells by adenovirus. Scale bar = 200 μm. (B) Analysis of the ability of Hakai-overexpressing BEL7402 cells by adenovirus to form colonies. Data are presented as Mean ± SEM from three independent experiments (**p < 0.01, ***p < 0.001). (JPG 146 kb

Additional file 2: of Ajuba inhibits hepatocellular carcinoma cell growth via targeting of β-catenin and YAP signaling and is regulated by E3 ligase Hakai through neddylation

Figure S2. Ajuba was co-localized with Hakai in HepG2 cells. (A) HepG2 cells were co-transfected with Myc-Ajuba or Myc-Vector and GFP-Hakai for 24 h. Cells were analyzed for GFP-Hakai/Myc-Ajuba co-localization, Scale bar = 25 μm. (JPG 97 kb

Additional file 1: of Ajuba inhibits hepatocellular carcinoma cell growth via targeting of β-catenin and YAP signaling and is regulated by E3 ligase Hakai through neddylation

Figure S1. The regulation of β-Catenin and cell growth in HCC cells. (A, B) HCC cells were transfected with specific siRNAs to silence GSK3β protein in Ajuba-depleted HCC cell lines. The expression of GSK3β, Ajuba, CyclinD1 and GAPDH were tested by immunoblot assay (A). β-Catenin translocation were tested by confocal assay, Scale bar = 25 μm (B). (C, D) HCC cells were transfected with specific siRNAs to silence β-Catenin protein in Ajuba-depleted HCC cell lines. The expression of β-Catenin, Ajuba, CyclinD1 and GAPDH were tested by immunoblot assay (C). Cell growth was tested by colony formation (D). (E) HCC cells were transfected with specific siRNAs to silence YAP protein in Ajuba-depleted HCC cell lines. Cell growth was tested by colony formation. Data are presented as Mean ± SEM from three independent experiments (***p < 0.001). (JPG 515 kb

Additional file 5: of Ajuba inhibits hepatocellular carcinoma cell growth via targeting of β-catenin and YAP signaling and is regulated by E3 ligase Hakai through neddylation

Figure S5. Ajuba knockdown-mediated β-catenin translocation into nucleus is not dependent on Hakai. (A, B) HepG2 cells were transfected with specific siRNAs to silence Ajuba protein in Hakai-depleted HepG2 cells. β-catenin translocation were tested by confocal assay, Scale bar = 25 μm (A). The expression of Ajuba, Hakai and β-catenin were tested by immunoblot assay, GAPDH was used as a loading control (B). (C) Immunoblot analysis of Ajuba and Hakai in BEL7402 and HepG2 cell lysis. GAPDH was used as a loading control. (JPG 617 kb