Journal: Cell Death Discovery

Article Title: MDMX reprograms glycolysis of hepatocellular carcinoma via 14-3-3γ/FOXO1

doi: 10.1038/s41420-025-02804-2

Figure Lengend Snippet: A In Huh7 cells with overexpression of MDMX, mRNA level of RPIA, PKM, HK2, PCK1, AQP9, and G6PC were detected by q-PCR. B In Huh7 cells with overexpression or knockdown of MDMX, the protein level of PCK1 and RPIA were measured by Western blot. C In Huh7 cells with overexpression of MDMX, mRNA level of FOXO1 was detected by q-PCR. D In Huh7 cells with overexpression or knockdown of MDMX, the protein level of FOXO1 were measured by Western blot. E ChIP-qPCR was conducted to analyze the combination of FOXO1 and the promoter of PCK1 in Huh7 cells with MDMX overexpression. F TCGA analysis of FOXO1 expression level in HCC tissues ( n = 50) compared with adjacent tissues ( n = 374) and the relationship to patient’s overall survival. G , H Immunohistochemical analysis of FOXO1 expression conducted on tumor samples from 48 patients with HCC alongside corresponding normal liver tissues. I CCK-8 and colony formation assay were conducted to study the effect of FOXO1 on cell growth. * P < 0.05, ** P < 0.01, *** P < 0.001. A – D , F , and H , I T -test was used for statistical analysis.

Article Snippet: Sections were then incubated with following primary antibody at 4 °C overnight: PCK1 (1:300, 16754-1-AP, Proteintech, China), RPIA (1:100, 13010-1-AP, Proteintech, China), FOXO1 (1:300, 18592-1-AP, Proteintech, China), and MDMX (1:500, A300-287A, Bethyl Laboratories, USA).

Techniques: Over Expression, Knockdown, Western Blot, ChIP-qPCR, Expressing, Immunohistochemical staining, CCK-8 Assay, Colony Assay

Journal: Cell Death Discovery

Article Title: MDMX reprograms glycolysis of hepatocellular carcinoma via 14-3-3γ/FOXO1

doi: 10.1038/s41420-025-02804-2

Figure Lengend Snippet: A Correlation analysis of MDMX and FOXO1 expression using immunohistochemical scoring ( n = 48). B , C mRNA and protein expression levels of PCK1 and RPIA after FOXO1 overexpression in Huh7 cells. D Glucose uptake, ATP levels, and lactate production were examined after FOXO1 overexpression in Huh7 cells. E Immunofluorescence images showed FOXO1 downregulation after MDMX overexpression. F Protein level of FOXO1 was measured by Western blot in Huh7 cells with overexpression of MDMX treated with 10 µM CHX. * P < 0.05, ** P < 0.01, *** P < 0.001. B–D , F T -test was used for statistical analysis.

Article Snippet: Sections were then incubated with following primary antibody at 4 °C overnight: PCK1 (1:300, 16754-1-AP, Proteintech, China), RPIA (1:100, 13010-1-AP, Proteintech, China), FOXO1 (1:300, 18592-1-AP, Proteintech, China), and MDMX (1:500, A300-287A, Bethyl Laboratories, USA).

Techniques: Expressing, Immunohistochemical staining, Over Expression, Immunofluorescence, Western Blot

Journal: Cell Death Discovery

Article Title: MDMX reprograms glycolysis of hepatocellular carcinoma via 14-3-3γ/FOXO1

doi: 10.1038/s41420-025-02804-2

Figure Lengend Snippet: A Colony formation and cell proliferation assays were conducted in Huh7 cells with MDMX knockdown and AS1842856 treatment. B , D Western blot showed the protein level of MDMX, FOXO1, PCK1, and RPIA after MDMX knockdown and treated with AS1842856 in Huh7 and Hep3B cells. C , E The level of glucose consumption, ATP level, and lactate production after MDMX knockdown and treated with AS1842856 in Huh7 and Hep3B cells. F , G Colony formation and cell proliferation assays were conducted in Huh7 cells with MDMX overexpression and 2-DG treatment. H Huh7 cells overexpressing MDMX were cultured with different glucose concentration medium (25, 12.5, 5 mM), colony formation ability was measured after 10 days cultural. * P < 0.05, ** P < 0.01, *** P < 0.001. A – G One-way ANOVA and H T -test were used for statistical analysis.

Article Snippet: Sections were then incubated with following primary antibody at 4 °C overnight: PCK1 (1:300, 16754-1-AP, Proteintech, China), RPIA (1:100, 13010-1-AP, Proteintech, China), FOXO1 (1:300, 18592-1-AP, Proteintech, China), and MDMX (1:500, A300-287A, Bethyl Laboratories, USA).

Techniques: Knockdown, Western Blot, Over Expression, Cell Culture, Concentration Assay

Journal: Cell Death Discovery

Article Title: MDMX reprograms glycolysis of hepatocellular carcinoma via 14-3-3γ/FOXO1

doi: 10.1038/s41420-025-02804-2

Figure Lengend Snippet: A Western blot showed the expression levels of FOXO1, PCK1, and RPIA in liver tissues from tissue specific expressing MDMX mice. B The levels of glucose uptake, ATP, and lactate production in the liver tissues were measured. C Immunohistochemistry showed the expression levels of FOXO1, PCK1, and RPIA in liver tissues from tissue specific expressing MDMX mice. D Mechanism diagram of how MDMX promotes the occurrence and development of liver cancer by promoting glycolysis via 14-3-3γ/FOXO1. * P < 0.05, ** P < 0.01. B T -test was used for statistical analysis.

Article Snippet: Sections were then incubated with following primary antibody at 4 °C overnight: PCK1 (1:300, 16754-1-AP, Proteintech, China), RPIA (1:100, 13010-1-AP, Proteintech, China), FOXO1 (1:300, 18592-1-AP, Proteintech, China), and MDMX (1:500, A300-287A, Bethyl Laboratories, USA).

Techniques: Western Blot, Expressing, Immunohistochemistry