anti chpf  (Proteintech)

Journal: Cells

Article Title: Targeting Chondroitin Sulfate Reduces Invasiveness of Glioma Cells by Suppressing CD44 and Integrin β1 Expression

doi: 10.3390/cells10123594

Figure Lengend Snippet: Correlation and modulation between CHSY1 and integrins in glioblastoma tissue and cells. ( A ) Gene expression of CHSY1 in comparison to related integrins. Data were collected from the cBioPortal database and analysed by Pearson’s correlation. ( B ) CHSY1, CHPF, and ITGB1 protein expressions in the glioblastoma cell lines GBM8401, Ln18, DBTRG, GBM8901, U251, A172, U118, and mouse glioblastoma cell line GL261. Each group was standardized with total protein. ( C ) ITGB1 protein expression upon overexpression or silencing of CHSY1 in glioma cells. Control siRNA (Ctr si) or CHSY1-siRNA (CHSY1 si). The red arrow indicates the mature form of ITGB1. ( D ) qPCR analysis of the gene expression of ITGB1 and CHSY1 in A172 and U118 cells with CHSY1 siRNA knockdown. Statistical significance, *** p < 0.001.

Article Snippet: In brief, equivalent amounts of cell lysates (30 μg) were applied to 8 % sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and blotted with antibodies against CD44 (Bioss, BSM51065M), CHSY1 (OriGene, Cat: TA315190, Rockville, MD, USA), CHPF (Proteintech, Cat: 239531AP, Chicago, IL, USA), ITGB1 (BD bioscience, Cat: 610467, Franklin Lakes, NJ, USA) and actin (Santa Cruz, sc-47778).

Techniques: Gene Expression, Comparison, Expressing, Over Expression, Control, Knockdown

Journal: Cancers

Article Title: CHPF Regulates the Aggressive Phenotypes of Hepatocellular Carcinoma Cells via the Modulation of the Decorin and TGF-β Pathways

doi: 10.3390/cancers13061261

Figure Lengend Snippet: CHPF expression in human hepatocellular carcinoma (HCC). ( A ) CHPF levels and quantification in non-tumor (peri-tumor) liver (N) and tumor tissue (T) of primary HCC tissues from twelve patients. ( B ) Immunohistochemistry of CHPF in a representative HCC tissue. Dot-like precipitations of CHPF (arrow) were observed mainly in peri-tumor tissues but rarely in stromal and tumor tissues. The scale bar indicates 50 μm. ( C ) Representative images of four grades of CHPF expression in primary HCC tissue (+0, negative; +1, <20%; +2, 20–50%; +3, >50%); n = 78. The scale bar indicates 150 μm. ( D ) Kaplan–Meier analysis of overall survival of HCC patients. The analyses were conducted according to the immunostaining of CHPF low expression (+0 and +1) and high expression (+2 and +3); p = 0.038.

Article Snippet: Mouse monoclonal antibody against CHPF (sc-376183) was purchased from Santa Cruz.

Techniques: Expressing, Immunohistochemistry, Immunostaining

Journal: Cancers

Article Title: CHPF Regulates the Aggressive Phenotypes of Hepatocellular Carcinoma Cells via the Modulation of the Decorin and TGF-β Pathways

doi: 10.3390/cancers13061261

Figure Lengend Snippet: Correlation of CHPF expression with clinicopathological features of hepatocellular carcinoma tissue.

Article Snippet: Mouse monoclonal antibody against CHPF (sc-376183) was purchased from Santa Cruz.

Techniques: Expressing

Journal: Cancers

Article Title: CHPF Regulates the Aggressive Phenotypes of Hepatocellular Carcinoma Cells via the Modulation of the Decorin and TGF-β Pathways

doi: 10.3390/cancers13061261

Figure Lengend Snippet: CHPF expression in HCC cells. ( A ) CHPF expression in seven HCC cell lines. Protein expression was analyzed by Western blotting, and total protein was used as an internal loading control. ( B ) Stable CHPF overexpression in Hepa1-6 and HA22T cells. Transient CHPF knockdown in HA59T cells 48 and 72 h after siRNA transfection. Protein expression was analyzed by Western blotting, and β-actin (ACTB) was used as an internal control. ( C ) Surface CS56 antibody staining (an anti-chondroitin sulfate antibody) on Hepa1-6, HA22T, and HA59T transfectants was analyzed by flow cytometry with anti-mouse IgM-FITC. Nonspecific mouse IgM was used as an isotype control (Iso). * p < 0.05, *** p < 0.001. ( D ) Fluorescence microscopy analysis of CS56 (green); DAPI (blue) indicated the position of the nucleus. The scale bar indicates 50 μm. The cells were fixed with paraformaldehyde without permeabilization.

Article Snippet: Mouse monoclonal antibody against CHPF (sc-376183) was purchased from Santa Cruz.

Techniques: Expressing, Western Blot, Control, Over Expression, Knockdown, Transfection, Staining, Flow Cytometry, Fluorescence, Microscopy

Journal: Cancers

Article Title: CHPF Regulates the Aggressive Phenotypes of Hepatocellular Carcinoma Cells via the Modulation of the Decorin and TGF-β Pathways

doi: 10.3390/cancers13061261

Figure Lengend Snippet: Effects of CHPF on HCC cell growth in vitro and in vivo. ( A ) CHPF modulated cell viability in vitro. Viability of Hepa1-6, HA22T, and HA59T cells was measured using CCK8 assays at indicted time points. Relative fold changes are shown. Data are presented as the means from three independent experiments ± standard deviations; ** p < 0.01. ( B ) CHPF overexpression suppressed colony formation in Hepa1-6 and HA22T cells. The data are presented as the means ± standard deviations; ** p < 0.01. Representative images are shown on the right. ( C ) Effects of CHPF on HCC tumor growth in mouse models. Hepa1-6 control transfectants and an equal number of CHPF transfectants were subcutaneously administered to the right flank ( n = 5, each group). Size and mass of the excised tumors were recorded; shown are the means ± standard errors of the means; * p < 0.05. The scale bar indicates 10 mm. Immunostaining of CHPF in tumor sections (lower right; the scale bar indicates 100 μm). ( D ) CHPF suppressed cell proliferation in Hepa1-6 tumors. Proliferation of tumor cells was evaluated by immunofluorescence staining for Ki67, and representative images of tumor section are shown (bottom). The data are presented as the means from five fields of each section ± standard deviations; *** p < 0.001. The scale bar indicates 100 μm.

Article Snippet: Mouse monoclonal antibody against CHPF (sc-376183) was purchased from Santa Cruz.

Techniques: In Vitro, In Vivo, Over Expression, Control, Immunostaining, Immunofluorescence, Staining

Journal: Cancers

Article Title: CHPF Regulates the Aggressive Phenotypes of Hepatocellular Carcinoma Cells via the Modulation of the Decorin and TGF-β Pathways

doi: 10.3390/cancers13061261

Figure Lengend Snippet: Influence of CHPF on the migration, invasion, and metastasis of HCC cells. ( A ) CHPF regulated transwell cell migration and ( B ) Matrigel invasion. CHPF overexpression significantly suppressed the migration and invasion abilities of Hepa1-6 and HA22T cells, and CHPF knockdown increased the migration and invasion abilities of HA59T cells. The data presented as the means from three independent experiments ± standard deviations. Representative images are shown at the bottom; * p < 0.05; ** p < 0.01; *** p < 0.001. ( C ) Immunostaining of ZO-1 (green) and F-actin (red) in HA59T cells. Amplified images are shown on the right. Note that ZO-1 is expressed on the plasma membrane of two neighboring control cells (indicated by arrows); CHPF knockdown increased the intracellular distribution of ZO-1 (indicated by arrowheads). The scale bar indicates 20 μm. The percentage of plasma membrane-associated ZO-1 positive cells was shown at right. *** p < 0.001. ( D ) CHPF suppressed lung metastasis. The number of tumors was decreased in the CHPF-overexpressing group. Representative images are shown at the bottom. Representative images of the hematoxylin and eosin staining and immunohistochemical staining for CHPF and Ki67 in the lung sections are shown on the right; ** p < 0.01.

Article Snippet: Mouse monoclonal antibody against CHPF (sc-376183) was purchased from Santa Cruz.

Techniques: Migration, Over Expression, Knockdown, Immunostaining, Amplification, Clinical Proteomics, Membrane, Control, Staining, Immunohistochemical staining

Journal: Cancers

Article Title: CHPF Regulates the Aggressive Phenotypes of Hepatocellular Carcinoma Cells via the Modulation of the Decorin and TGF-β Pathways

doi: 10.3390/cancers13061261

Figure Lengend Snippet: TGF-β signaling modulates CHPF-mediated malignant phenotypes. ( A ) Hepa1-6 and HA59T transfectants were starved for 3 h and then treated with (+) or without (−) TGF-β (20 ng/mL) for 15 and 30 min. Cell lysates (20 μg) were analyzed by Western blotting with various antibodies, as indicated. ( B ) Effects of LY364947, a TGF-β inhibitor, on CHPF-mediated cell viability. HA59T transfectants were treated with a solvent (0.02% DMSO) or LY364947 (0.5 μM) and then analyzed by CCK8 assay. * p < 0.05, ** p < 0.01. ( C ) Effects of LY364947 on CHPF-mediated migration (left) and invasion (right). HA59T transfectants were treated with a solvent (0.1% DMSO) or LY364947 (2.5 μM) and then analyzed by transwell migration and invasion assays. The data are shown as the means ± standard deviations. ** p < 0.01.

Article Snippet: Mouse monoclonal antibody against CHPF (sc-376183) was purchased from Santa Cruz.

Techniques: Western Blot, Solvent, CCK-8 Assay, Migration

Journal: Cancers

Article Title: CHPF Regulates the Aggressive Phenotypes of Hepatocellular Carcinoma Cells via the Modulation of the Decorin and TGF-β Pathways

doi: 10.3390/cancers13061261

Figure Lengend Snippet: Decorin is modified by CHPF and is involved in malignant phenotypes in HCC cells. ( A ) CHPF modified the CS chains on decorin (DCN). The molecular weight of glycanated decorin was decreased after CHPF knockdown in HA59T cells. Glycanated decorin was mostly shifted to the size of core protein after 2 h of digestion with chondroitinase ABC (ChABC). These results showed no difference with regard to heparinase II treatments. Blotting of ACTB was used as an internal control. ( B ) Immunostaining of DCN in controls and CHPF-silenced cells. Cells with paraformaldehyde (PFA) fixation are shown at the top. Images of PFA fixation, followed by triton X100 permeabilization, are shown at the bottom. The scale bar indicates 50 μm. ( C ) Secreted DCN in HA59T culture medium (1 × 10 6 of cells in 1 mL serum free medium for 24 h). Blotting of ACTB from attached cell lysate was used as a loading control. ( D ) Silence of DCN enhanced TGF-β signaling. Effects of DCN silencing on CHPF-mediated cell viability ( E ) and cell invasion ( F ). The HA59T cells were transfected with non-targeting control siRNAs (Ctr si), CHPF si, and/or DCN specific siRNAs (DCN si). Cells were subjected to CCK8 assay or transwell invasion assay. Data are expressed as the mean ± SD from three independent experiments. ** p < 0.01. The representative images are shown on the right. ( G ) Western blots of DCN and CHPF in 16 primary HCC tissues. ACTB was used as a loading control. ( H ) Correlations between CHPF protein expression and glycanated DCN in HCC tissues; Pearson r 2 = 0.5257; p = 0.0015.

Article Snippet: Mouse monoclonal antibody against CHPF (sc-376183) was purchased from Santa Cruz.

Techniques: Modification, Molecular Weight, Knockdown, Control, Immunostaining, Transfection, CCK-8 Assay, Transwell Invasion Assay, Western Blot, Expressing