cstf2 rabbit pabs antibody  (ABclonal Biotechnology)

Journal: Frontiers in Oncology

Article Title: CSTF2 Promotes Hepatocarcinogenesis and Hepatocellular Carcinoma Progression via Aerobic Glycolysis

doi: 10.3389/fonc.2022.897804

Figure Lengend Snippet: The expression levels of CSTF2 in human cancers. (A) The expression levels of CSTF2 in different tumor tissues compared to normal tissues in The Cancer Genome Atlas (TCGA) database. (B) The expression of CSTF2 in tumor tissues and matched TCGA normal in TCGA database and GTEx data by GEPIA analysis. * ρ < 0.05, ** ρ < 0.01, *** ρ < 0.001 compared to normal tissues.

Article Snippet: The polyvinylidene fluoride (PVDF) membranes were blocked with non-fat milk and incubated with the primary antibodies against β-actin mouse antibody (1:1,000, ABclonal, China) and CSTF2 rabbit pAbs (1:1,000, ABclonal, Wuhan, China) overnight at 4°C, washed with TBST and then incubated with the corresponding HRP-conjugated secondary antibodies (Thermo Fisher Science, USA) for 1 h at room temperature (RT) and washed with a Tris-buffered saline with Tween-20 (TBST) buffer.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: CSTF2 Promotes Hepatocarcinogenesis and Hepatocellular Carcinoma Progression via Aerobic Glycolysis

doi: 10.3389/fonc.2022.897804

Figure Lengend Snippet: CSTF2 is highly expressed in HCC. (A) Analysis of CSTF2 mRNA expression levels in TCGA, International Cancer Genome Consortium (ICGC), and Chinese patients with hepatitis B virus (CHCC-HBV) databases (B) Representative images of the immunohistochemistry staining of CSTF2 in hepatocellular carcinoma (HCC) and adjacent non-tumor tissues from different HCC patients (tumor indicates HCC tissue; normal indicates adjacent normal tissue). (C) Quantitative analysis of sample numbers in different CSTF2 expression levels in 48 pairs of HCC tissues. (D) Western blot analysis of CSTF2 protein levels in non-tumorigenic LO2 liver cells, hepatocellular carcinoma cell lines Huh7, MHCC-97H, and Hep3B (left panel) and qualification of the relative intensity of CSTF2 protein levels (right panel). The levels of β-actin were used as an internal control. ** ρ < 0.01, *** ρ < 0.001.

Article Snippet: The polyvinylidene fluoride (PVDF) membranes were blocked with non-fat milk and incubated with the primary antibodies against β-actin mouse antibody (1:1,000, ABclonal, China) and CSTF2 rabbit pAbs (1:1,000, ABclonal, Wuhan, China) overnight at 4°C, washed with TBST and then incubated with the corresponding HRP-conjugated secondary antibodies (Thermo Fisher Science, USA) for 1 h at room temperature (RT) and washed with a Tris-buffered saline with Tween-20 (TBST) buffer.

Techniques: Expressing, Virus, Immunohistochemistry, Staining, Western Blot, Control

Journal: Frontiers in Oncology

Article Title: CSTF2 Promotes Hepatocarcinogenesis and Hepatocellular Carcinoma Progression via Aerobic Glycolysis

doi: 10.3389/fonc.2022.897804

Figure Lengend Snippet: The mRNA levels of CSTF2 were correlated with the advanced clinical stage in HCC in different databases. (A-F) The relationship between CSTF2 expression levels with a histological grade, a clinical stage in TCGA databases, ICGC databases, CHCC-HBV databases, GSE14520 databases, and GSE76427 databases and in GSE36376 databases. ** ρ < 0.01, *** p < 0.001 compared to normal.

Article Snippet: The polyvinylidene fluoride (PVDF) membranes were blocked with non-fat milk and incubated with the primary antibodies against β-actin mouse antibody (1:1,000, ABclonal, China) and CSTF2 rabbit pAbs (1:1,000, ABclonal, Wuhan, China) overnight at 4°C, washed with TBST and then incubated with the corresponding HRP-conjugated secondary antibodies (Thermo Fisher Science, USA) for 1 h at room temperature (RT) and washed with a Tris-buffered saline with Tween-20 (TBST) buffer.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: CSTF2 Promotes Hepatocarcinogenesis and Hepatocellular Carcinoma Progression via Aerobic Glycolysis

doi: 10.3389/fonc.2022.897804

Figure Lengend Snippet: CSTF2 higher expression is associated with poor prognosis in HCC. (A-D) Overall survival and relapse-free survival in TCGA-LIHC, CHCC-HBV, GSE14520, and ICGC databases (E) Univariate and (F) multivariate Cox analyses of CSTF2 expression at different TNM stages, ages, and genders.

Article Snippet: The polyvinylidene fluoride (PVDF) membranes were blocked with non-fat milk and incubated with the primary antibodies against β-actin mouse antibody (1:1,000, ABclonal, China) and CSTF2 rabbit pAbs (1:1,000, ABclonal, Wuhan, China) overnight at 4°C, washed with TBST and then incubated with the corresponding HRP-conjugated secondary antibodies (Thermo Fisher Science, USA) for 1 h at room temperature (RT) and washed with a Tris-buffered saline with Tween-20 (TBST) buffer.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: CSTF2 Promotes Hepatocarcinogenesis and Hepatocellular Carcinoma Progression via Aerobic Glycolysis

doi: 10.3389/fonc.2022.897804

Figure Lengend Snippet: Knockout of CSTF2 inhibited the proliferation, migration, and invasion of HCC cells in vitro . (A) The stable CSTF2 knockout Huh7 and MHCC-97H cells were determined by Western blot (upper panel) and the qualification of the relative intensity of CSTF2 protein levels (bottom panel). The levels of β-actin were used as an internal control. (B) The cell proliferation ability of CSTF2 knockout Huh7 and MHCC-97H cells was measured by the CCK-8 assay. (C) Colony-forming assay of CSTF2 knockout Huh7 cells and MHCC-97H cells (left panel) and the qualification of the number of colonies in these cells (right panel). (D) The migration and invasion capacity were evaluated in CSTF2 knockout Huh7 cells and MHCC-97H cells by the Transwell assay (upper panel) and the qualification of migrated cells and invading cells per field (bottom panel). Scale bar, 50 μm. sgCSTF2 represents CSTF2-sgRNA. An empty vector was used as a control of CSTF2-sgRNA. * ρ < 0.05, ** p < 0.01, *** p < 0.001 compared to vector control.

Article Snippet: The polyvinylidene fluoride (PVDF) membranes were blocked with non-fat milk and incubated with the primary antibodies against β-actin mouse antibody (1:1,000, ABclonal, China) and CSTF2 rabbit pAbs (1:1,000, ABclonal, Wuhan, China) overnight at 4°C, washed with TBST and then incubated with the corresponding HRP-conjugated secondary antibodies (Thermo Fisher Science, USA) for 1 h at room temperature (RT) and washed with a Tris-buffered saline with Tween-20 (TBST) buffer.

Techniques: Knock-Out, Migration, In Vitro, Western Blot, Control, CCK-8 Assay, Transwell Assay, Plasmid Preparation

Journal: Frontiers in Oncology

Article Title: CSTF2 Promotes Hepatocarcinogenesis and Hepatocellular Carcinoma Progression via Aerobic Glycolysis

doi: 10.3389/fonc.2022.897804

Figure Lengend Snippet: Overexpression of CSTF2 promoted the cell proliferation, migration, and invasion of HCC cells in vitro . (A) The stable overexpression of CSTF2 in Hep3B cells was determined by Western blot (left panel) and the qualification of the relative intensity of CSTF2 protein levels (right panel). (B) CCK-8 assay of the cell proliferation rate of the overexpression of CSTF2 Hep3B cells. (C) Colony-forming assay of the overexpression of CSTF2 Hep3B cells (left panel) and the qualification of the number of colonies formed (right panel). (D) The migration and invasion were measured in the overexpression of CSTF2 Hep3B cells by the Transwell assay (left panel) and the qualification of migrated cells and invading cells per field (right panel). CSTF2 represents the overexpression of CSTF2. An empty vector was used as control. Scale bar, 50 μm. * ρ < 0.05, ** ρ < 0.01, *** ρ < 0.001 compared to the vector.

Article Snippet: The polyvinylidene fluoride (PVDF) membranes were blocked with non-fat milk and incubated with the primary antibodies against β-actin mouse antibody (1:1,000, ABclonal, China) and CSTF2 rabbit pAbs (1:1,000, ABclonal, Wuhan, China) overnight at 4°C, washed with TBST and then incubated with the corresponding HRP-conjugated secondary antibodies (Thermo Fisher Science, USA) for 1 h at room temperature (RT) and washed with a Tris-buffered saline with Tween-20 (TBST) buffer.

Techniques: Over Expression, Migration, In Vitro, Western Blot, CCK-8 Assay, Transwell Assay, Plasmid Preparation, Control

Journal: Frontiers in Oncology

Article Title: CSTF2 Promotes Hepatocarcinogenesis and Hepatocellular Carcinoma Progression via Aerobic Glycolysis

doi: 10.3389/fonc.2022.897804

Figure Lengend Snippet: Knockout of CSTF2 inhibited the tumorigenesis and progression of HCC. (A) In vivo tumor formation of CSTF2 sgRNA Huh7 and vector control Huh7. Tumorigenesis experiments were performed by subcutaneously injecting Huh7 cells into BALB/c nude mice. (B) Strategy for evaluating the hepatocarcinogenesis function of CSTF2 in immunocompetent C57BL/6 mice. Schematic representation of the hydrodynamic tail vein injection HCC model. Plasmids pT3-EF1A-MYC-IRES-luc, px330-p53 (p53-sgRNA), and CMV-SB13 transposase were delivered together with either CSTF2 sgRNA (sgCSTF2) or the control vector for HCC induction. (C) Representative images of dissected livers in CSTF2 sgRNA and the vector control group. (D) The number of tumor nodules in mice injected with sgCSTF2 was significantly lower than that in the control group. (E) Qualification of the liver weight, and the ratio of liver weight to body weight of these tumor nodules in CSTF2 sgRNA or control groups. At least 6 mice were used in every group. (F) Immunohistochemical staining formed tumors in the liver tissues using CSTF2 and Ki67 antibodies. Scale bar, 100 μm. sgCSTF2 represents CSTF2-sgRNA. The empty vector of sgRNA was used as control. * ρ < 0.05, ** ρ < 0.01 compared to control.

Article Snippet: The polyvinylidene fluoride (PVDF) membranes were blocked with non-fat milk and incubated with the primary antibodies against β-actin mouse antibody (1:1,000, ABclonal, China) and CSTF2 rabbit pAbs (1:1,000, ABclonal, Wuhan, China) overnight at 4°C, washed with TBST and then incubated with the corresponding HRP-conjugated secondary antibodies (Thermo Fisher Science, USA) for 1 h at room temperature (RT) and washed with a Tris-buffered saline with Tween-20 (TBST) buffer.

Techniques: Knock-Out, In Vivo, Plasmid Preparation, Control, Injection, Immunohistochemical staining, Staining

Journal: Frontiers in Oncology

Article Title: CSTF2 Promotes Hepatocarcinogenesis and Hepatocellular Carcinoma Progression via Aerobic Glycolysis

doi: 10.3389/fonc.2022.897804

Figure Lengend Snippet: Functional enrichment analysis of CSTF2 in the TCGA cohort. (A) GO classification analysis of gene changes in the biological process. (B) KEGG pathway enrichment analysis of upregulated pathways (C, D) GSEA analysis of activated pathways and hallmarks of CSTF2 related in HCC based on the TCGA database.

Article Snippet: The polyvinylidene fluoride (PVDF) membranes were blocked with non-fat milk and incubated with the primary antibodies against β-actin mouse antibody (1:1,000, ABclonal, China) and CSTF2 rabbit pAbs (1:1,000, ABclonal, Wuhan, China) overnight at 4°C, washed with TBST and then incubated with the corresponding HRP-conjugated secondary antibodies (Thermo Fisher Science, USA) for 1 h at room temperature (RT) and washed with a Tris-buffered saline with Tween-20 (TBST) buffer.

Techniques: Functional Assay

Journal: Frontiers in Oncology

Article Title: CSTF2 Promotes Hepatocarcinogenesis and Hepatocellular Carcinoma Progression via Aerobic Glycolysis

doi: 10.3389/fonc.2022.897804

Figure Lengend Snippet: High expression of CSTF2 enhances aerobic glycolysis in HCC. (A) Heat map of the cluster analysis of the relationship between the CSTF2 expression and glycolysis-related enzymes and glucose transporters. (B) The correlation analysis of CSTF2 expression and glycolysis-related enzymes and glucose transporters. (C-D) The relative expression levels of CSTF2, SLC2A1, HK2, LDHA, PKM, PFKFB3, and PFKM detected by quantitative RT-PCR in Huh7 cells and MHCC-97H cells. (E–F) The protein levels of HK2 in Huh7 cells and MHCC-97H cells determined by Western blot; the levels of β-actin were used as an internal control. (G) 3’ RACE HK2 fragments were amplified in Huh7 cells and detected by agarose gel electrophoresis. (H–J) The glycolytic function of HCC cells was measured by the extracellular acidification rate (EACR), relative glucose uptake, and lactate production in CSTF2 knockout Huh7 and MHCC-97H cells. (K-L) The glycolytic function of HCC cells was measured by EACR, the relative glucose uptake, and lactate production in CSTF2-overexpression Hep3B cells. sgCSTF2 represents CSTF2 sgRNA, CSTF2 represents CSTF2 overexpression, and the vector was used as control. Data were presented as the mean ± SEM. ** ρ < 0.01, *** ρ < 0.001 compared to the vector.

Article Snippet: The polyvinylidene fluoride (PVDF) membranes were blocked with non-fat milk and incubated with the primary antibodies against β-actin mouse antibody (1:1,000, ABclonal, China) and CSTF2 rabbit pAbs (1:1,000, ABclonal, Wuhan, China) overnight at 4°C, washed with TBST and then incubated with the corresponding HRP-conjugated secondary antibodies (Thermo Fisher Science, USA) for 1 h at room temperature (RT) and washed with a Tris-buffered saline with Tween-20 (TBST) buffer.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Amplification, Agarose Gel Electrophoresis, Knock-Out, Over Expression, Plasmid Preparation