Journal: Cell Biochemistry and Function

Article Title: Zinc Finger Protein 143 Bidirectionally Regulates the Proliferation of Hepatocytes and Hepatocellular Carcinoma Cells via c‐Myc

doi: 10.1002/cbf.70320

Figure Lengend Snippet: The effects of ZNF143 knockdown on the expression of cell proliferation and cell cycle‐related genes in AML12 cells. (A) qRT‐PCR analysis of ZNF143 mRNA levels. (B) CCK‐8 assay evaluating AML12 cell viability at 48 and 72 h after ZNF143 knockdown. (C) MTT assay evaluating AML12 cell viability at 48 and 72 h after ZNF143 knockdown. (D) EdU fluorescence imaging; 4′,6‐diamidino‐2‐phenylindole (DAPI) was used for nuclear staining, and EdU incorporation was used to label proliferating cells. (E) Analysis of EdU‐positive cell rates. (F) qRT‐PCR analysis of the mRNA levels of proliferation and cell cycle‐related genes. (G) Western blot analysis of the protein levels of proliferation and cell cycle‐related genes; GAPDH was used as the loading control. (H) Grayscale analysis of the bands in (G). All data are presented as mean ± SEM, n = 3 (three biological replicates), statistical significance was determined by a two‐sided Student's t test, with the control group (set as the reference baseline) used for pairwise comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mouse hepatocyte AML12 cells (RRID: CRL‐2254, ATCC, Manassas, VA, USA) were cultured in Dulbecco's modified Eagle's medium (DMEM)/F‐12 medium (Hyclone) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY), 1x ITS‐G and 100 nM dexamethasone, HEK293T cells (RRID: CRL‐3216, ATCC, Manassas, VA, USA) were cultured using DMEM containing 10% FBS.

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Analysis, CCK-8 Assay, MTT Assay, Fluorescence, Imaging, Staining, EdU Assay, Western Blot, Control

Journal: Cell Biochemistry and Function

Article Title: Zinc Finger Protein 143 Bidirectionally Regulates the Proliferation of Hepatocytes and Hepatocellular Carcinoma Cells via c‐Myc

doi: 10.1002/cbf.70320

Figure Lengend Snippet: ZNF143 regulates the expression of c‐Myc in AML12 cells. (A) qRT‐PCR analysis of c‐Myc mRNA levels in AML12 cells upon ZNF143 knockdown. (B) Western blot analysis of c‐Myc protein levels in AML12 cells upon ZNF143 knockdown; GAPDH was used as the loading control. (C) Grayscale analysis of the bands in (B). (D) qRT‐PCR analysis of c‐Myc mRNA levels in AML12 cells upon ZNF143 OE; GAPDH was used as the loading control. (E) Western blot analysis of c‐Myc protein levels in AML12 cells upon ZNF143 OE; GAPDH was used as the loading control. (F) Grayscale analysis of the bands in (E) All data are presented as mean ± SEM, n = 3 (three biological replicates), statistical significance was determined by a two‐sided Student's t test, with the control group (set as the reference baseline) used for pairwise comparisons. * p < 0.05, *** p < 0.001.

Article Snippet: Mouse hepatocyte AML12 cells (RRID: CRL‐2254, ATCC, Manassas, VA, USA) were cultured in Dulbecco's modified Eagle's medium (DMEM)/F‐12 medium (Hyclone) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY), 1x ITS‐G and 100 nM dexamethasone, HEK293T cells (RRID: CRL‐3216, ATCC, Manassas, VA, USA) were cultured using DMEM containing 10% FBS.

Techniques: Expressing, Quantitative RT-PCR, Analysis, Knockdown, Western Blot, Control

Journal: Cell Biochemistry and Function

Article Title: Zinc Finger Protein 143 Bidirectionally Regulates the Proliferation of Hepatocytes and Hepatocellular Carcinoma Cells via c‐Myc

doi: 10.1002/cbf.70320

Figure Lengend Snippet: The effects of OE of ZNF143, c‐Myc, and their co‐OE on cell viability and proliferation in AML12 cells. (A) MTT assay evaluating AML12 cell viability at 48 and 72 h after ZNF143 OE. (B) MTT assay evaluating AML12 cell viability at 48 and 72 h after c‐Myc OE. (C) MTT assay evaluating AML12 cell viability at 48 and 72 h after co‐OE of ZNF143 and c‐Myc. (D) CCK‐8 assay evaluating AML12 cell viability at 48 and 72 h after ZNF143 OE. (E) CCK‐8 assay evaluating AML12 cell viability at 48 and 72 h after c‐Myc OE. (F) CCK‐8 assay evaluating AML12 cell viability at 48 and 72 h after co‐OE of ZNF143 and c‐Myc. (G) EdU fluorescence imaging; DAPI was used for nuclear staining, and EdU incorporation was used to label proliferating cells. (H) Analysis of EdU‐positive cell rates. (I) qRT‐PCR analysis of ZNF143 and c‐Myc mRNA levels. (J) qRT‐PCR analysis of the mRNA levels of cell proliferation and cell cycle‐related genes. All data are presented as mean ± SEM, n = 3 (three biological replicates), statistical significance was determined by a two‐sided Student's t test, with the control group (set as the reference baseline) used for pairwise comparisons. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mouse hepatocyte AML12 cells (RRID: CRL‐2254, ATCC, Manassas, VA, USA) were cultured in Dulbecco's modified Eagle's medium (DMEM)/F‐12 medium (Hyclone) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY), 1x ITS‐G and 100 nM dexamethasone, HEK293T cells (RRID: CRL‐3216, ATCC, Manassas, VA, USA) were cultured using DMEM containing 10% FBS.

Techniques: MTT Assay, CCK-8 Assay, Fluorescence, Imaging, Staining, EdU Assay, Analysis, Quantitative RT-PCR, Control

Journal: Cell Biochemistry and Function

Article Title: Zinc Finger Protein 143 Bidirectionally Regulates the Proliferation of Hepatocytes and Hepatocellular Carcinoma Cells via c‐Myc

doi: 10.1002/cbf.70320

Figure Lengend Snippet: A proposed model illustrating the role of ZNF143 and c‑Myc in the proliferation of normal (AML12) and hepatoma (Huh7) cell lines. The left and right panels represent AML12 and Huh7 cells, respectively. In AML12 cells, ZNF143 inhibits c‐Myc expression and suppresses cell cycle progression and proliferation. In Huh7 cells, ZNF143 promotes cell cycle progression and proliferation. Green circles represent ZNF143; red circles represent c‐Myc. Arrows indicate promotion, and blunt‐ended lines indicate inhibition. The figure was created with the assistance of Adobe Illustrator (24.0.3).

Article Snippet: Mouse hepatocyte AML12 cells (RRID: CRL‐2254, ATCC, Manassas, VA, USA) were cultured in Dulbecco's modified Eagle's medium (DMEM)/F‐12 medium (Hyclone) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY), 1x ITS‐G and 100 nM dexamethasone, HEK293T cells (RRID: CRL‐3216, ATCC, Manassas, VA, USA) were cultured using DMEM containing 10% FBS.

Techniques: Expressing, Inhibition

Journal: Journal of Inflammation Research

Article Title: Vitamin B12 Alleviates Hepatic Ischemia–Reperfusion Injury with Restoration of Propanoate Metabolism and Mitochondrial Function

doi: 10.2147/JIR.S616890

Figure Lengend Snippet: VB12 can alleviate H/R-induced damage in AML12 cells. ( A ) Schematic diagram of the experimental procedure. ( B ) Alamar Blue assay to detect cell viability of hepatocytes after H/R treatment with different concentrations of vitamin B12 (0.3–300μM). ( C ) Flow cytometry assay to detect the proportion of Annexin V + apoptotic cells and Ki67 + proliferating cells, and the MFI of IL-6 and IL-1β. ( D ) qRT-PCR assay to detect the mRNA expression levels of Il6, Il1b, Tnf, Ccl2 and Cxcl2 . Data are expressed as mean ± SEM (n = 4 per group). Significance indicators, *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.

Article Snippet: AML12 cells were obtained from the American Type Culture Collection (ATCC; CRL-2254) and cultured in DMEM/F12 medium containing 10% fetal bovine serum (FBS) and supplemented with ITS and dexamethasone.

Techniques: Alamar Blue Assay, Flow Cytometry, Quantitative RT-PCR, Expressing