Journal: American Journal of Cancer Research

Article Title: Downregulation of Notch3 links TIMP3 inhibition to suppress aggressive phenotypes of pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Suppression of Notch3 reduced BxPC3 cell proliferation, migration and invasion. A. Western blots showing Notch1-4 protein expressions in human PC cell lines. B. The protein (left) and mRNA (right) levels of Notch3 were assessed in BxPC3 cells transfected with Notch3 siRNAs and siControl by Western blotting and QPCR. C-E. MTT, wound healing, and Transwell assays were performed to estimate the cell proliferation and motility of BxPC3 cells transfected with siNotch3 and siControl. F. Notch3 downstream targets were analyzed by QPCR in BxPC3 cells transfected with siNotch3 or siControl. Data are presented as the mean ± SD. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: Human pancreatic/duct cancer cell lines BxPC3 (RRID:CVCL_0186) and Panc-1 (RRID:CVCL_0480) were obtained from ATCC (American Type Culture Collection) and cultured in DMEM (Life Technologies, Inc., Carlsbad, USA) supplemented with 10% fetal bovine serum (FBS) (Life Technologies, Inc., Carlsbad, USA), 100 U/ml penicillin and 100 μg/ml streptomycin, 1% non-essential amino acid and 1% sodium pyruvate (Life Technologies, Inc., Carlsbad, USA).

Techniques: Migration, Western Blot, Transfection

Journal: American Journal of Cancer Research

Article Title: Downregulation of Notch3 links TIMP3 inhibition to suppress aggressive phenotypes of pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Knocking down Notch3 resulted in increased gemcitabine sensitivity in BxPC3 cells. A. Protein expression level of Notch3 in BxPC3 cells transfected with siNotch3 or siControl in the absence or presence of gemcitabine. B. MTT assay was performed to examine cell viability following gemcitabine treatment in siNotch3 and siControl cells. C. The migration and invasion assays were performed by Transwell in siNotch3 and siControl cells after gemcitabine (0.025 μM) treatment for 24 h. Data are presented as means ± SD. **P<0.01; ***P<0.001.

Article Snippet: Human pancreatic/duct cancer cell lines BxPC3 (RRID:CVCL_0186) and Panc-1 (RRID:CVCL_0480) were obtained from ATCC (American Type Culture Collection) and cultured in DMEM (Life Technologies, Inc., Carlsbad, USA) supplemented with 10% fetal bovine serum (FBS) (Life Technologies, Inc., Carlsbad, USA), 100 U/ml penicillin and 100 μg/ml streptomycin, 1% non-essential amino acid and 1% sodium pyruvate (Life Technologies, Inc., Carlsbad, USA).

Techniques: Expressing, Transfection, MTT Assay, Migration

Journal: American Journal of Cancer Research

Article Title: Downregulation of Notch3 links TIMP3 inhibition to suppress aggressive phenotypes of pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: TIMP3 was a key effector of the Notch3-raised aggressive phenotypes in BxPC3 cells. A. Venn diagrams comparing genes that were up or down-regulated in BxPC3 and Panc-1 cells transfected with siNotch3 or siControl. B. The protein expression level of TIMP3 was examined in siNotch3- and siControl-BxPC3 cells. C. The expression level of extracellular matrix TIMP3 was assessed in siNotch3-BxPC3 and siContrtol-BxPC3 cells by ELISA. D. ChIP assays were performed to determine the binding of Notch3 to the TIMP3 promoter in BxPC3 cells using an anti-Notch3 antibody. Isotype IgGs were used as a negative control. E. Western blot analysis of siNotch3 cells transfected with Flag-TIMP3 or vector alone. F and G. Cell proliferation, migration and invasion of siNotch3/vector and siNotch3/TIMP3 were estimated. H. The increase of TIMP3 mRNA in PAAD samples were found in the Oncomine database. I. A positive correlation between Notch3 mRNA and TIMP3 mRNA was found in the GEPIA cohort. Data are presented as the means ± SD. **P<0.01; ***P<0.001.

Article Snippet: Human pancreatic/duct cancer cell lines BxPC3 (RRID:CVCL_0186) and Panc-1 (RRID:CVCL_0480) were obtained from ATCC (American Type Culture Collection) and cultured in DMEM (Life Technologies, Inc., Carlsbad, USA) supplemented with 10% fetal bovine serum (FBS) (Life Technologies, Inc., Carlsbad, USA), 100 U/ml penicillin and 100 μg/ml streptomycin, 1% non-essential amino acid and 1% sodium pyruvate (Life Technologies, Inc., Carlsbad, USA).

Techniques: Transfection, Expressing, Enzyme-linked Immunosorbent Assay, Binding Assay, Negative Control, Western Blot, Plasmid Preparation, Migration

Journal: American Journal of Cancer Research

Article Title: Downregulation of Notch3 links TIMP3 inhibition to suppress aggressive phenotypes of pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: TIMP3 activation reversed siNotch3-increased sensitivity to gemcitabine in BxPC3 cells. A. Protein expression levels of Notch3 and TIMP3 in siNotch3 and siControl BxPC3 cells in the absence or presence of gemcitabine. B. Cell viability was accessed in siNotch3-BxPC3 cells transfected with vector or pCMV6-TIMP3 in a gemcitabine. C and D. Cell migratory and invasive abilities of indicated cells treated with gemcitabine. Data are presented as means ± SD. *P<0.05; **P<0.01; ***P<0.001.

Article Snippet: Human pancreatic/duct cancer cell lines BxPC3 (RRID:CVCL_0186) and Panc-1 (RRID:CVCL_0480) were obtained from ATCC (American Type Culture Collection) and cultured in DMEM (Life Technologies, Inc., Carlsbad, USA) supplemented with 10% fetal bovine serum (FBS) (Life Technologies, Inc., Carlsbad, USA), 100 U/ml penicillin and 100 μg/ml streptomycin, 1% non-essential amino acid and 1% sodium pyruvate (Life Technologies, Inc., Carlsbad, USA).

Techniques: Activation Assay, Expressing, Transfection, Plasmid Preparation