Protein Extraction and Western Blot Analysis

Protein extraction was conducted using RIPA Lysis Buffer (Beyotime Biotechnology, Shanghai, China) containing a protease inhibitor cocktail (Thermo Fisher Scientific, USA). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) assay was performed as in previous reports.^{52 (link)} Primary antibodies used in this study were β-actin (cat. 200068-8F10; ZEN-bioscience), H3 (cat. ASO71; ABclonal), Cyclin D1 (cat. 55506 T; CST), CDK4 (cat. 12790 T; CST), Cyclin A2 (cat. ab181591; Abcam), Cyclin E1 (cat. 20808 S; CST), CDK1 (cat. ab32094; Abcam), Cyclin B1(cat. 55004-1-AP; Proteintech), H3K9me1 (cat. A2358; ABclonal), H3K9me2 (cat. A2359; ABclonal), H3K9me3 (cat. ab176916; Abcam), MDIG (cat. 12214-1-AP; Proteintech), OTX2 (cat. 13497-1-AP; Proteintech), Myc (cat. 10828-1-AP; Proteintech), BRCA2 (cat. AF7817; Affinity), Junb (cat. 10486-1AP; Proteintech), E2F2 (cat. AF4100), ATAD5 (cat. Bs-7656R; Bioss). Western blots were quantified using ImageJ software (Version 1.51n, National Institutes of Health, Bethesda, MD, https://imagej.net/ij/index.html), as previously described.^{53 (link)} The original and uncropped films of Western blots were provided as Supplementary Figs. S19–S21.

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Du J., Liao W., Wang H., Hou G., Liao M., Xu L., Huang J., Yuan K., Chen X, & Zeng Y. (2023). MDIG-mediated H3K9me3 demethylation upregulates Myc by activating OTX2 and facilitates liver regeneration. Signal Transduction and Targeted Therapy, 8, 351.

Publication 2023

RIPA Lysis Buffer protease inhibitor cocktail β-actin Cyclin D1 Cyclin B1 H3K9me2 H3K9me3 BRCA2

Actin Antibodies Assay Atad5 Brca2 Buffer Cat 1 Cdk1 Cyclin a2 Cyclin b1 Cyclin d1 Cyclin e1 Figs Protease inhibitor Protein Ripa Sds page Western blots

Corresponding Organization :

Other organizations : Sichuan University, Jinan University, Sichuan Cancer Hospital, University of Electronic Science and Technology of China, West China Hospital of Sichuan University

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Variable analysis

independent variables

RIPA Lysis Buffer (Beyotime Biotechnology, Shanghai, China) containing a protease inhibitor cocktail (Thermo Fisher Scientific, USA)

dependent variables

Protein expression levels of the following proteins: β-actin, H3, Cyclin D1, CDK4, Cyclin A2, Cyclin E1, CDK1, Cyclin B1, H3K9me1, H3K9me2, H3K9me3, MDIG, OTX2, Myc, BRCA2, Junb, E2F2, ATAD5

control variables

SDS-PAGE assay was performed as in previous reports

controls

No positive or negative controls were explicitly mentioned in the information provided.

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