Journal: BMC Cancer

Article Title: Oncogenic potential of N-terminal deletion and S45Y mutant β-catenin in promoting hepatocellular carcinoma development in mice

doi: 10.1186/s12885-018-4870-z

Figure Lengend Snippet: β-catenin mutants cooperate with c-Met to promoter HCC development in mice. a Study design. b and c Liver weight and liver/body weight ratio among wild-type mice, c-Met/β-cateninS45Y mice, and c-Met/∆N90-β-catenin mice. Data are presented as mean ± standard deviation. Student’s t-test: **: P < 0.01, ***: P < 0.001; NS: Not significant

Article Snippet: After a blocking step with the 5% goat serum and Avidin-Biotin blocking kit (Vector Laboratories Inc., Burlingame, CA), the slides were incubated with the following primary antibodies: mouse anti-β-catenin (1:200; BD Transduction LaboratoriesTM, San Jose, CA), mouse anti-Glutamine Synthetase (1:500; BD Transduction LaboratoriesTM), β-catenin (D10A8) XP® Rabbit mAb (1:100; Cell Signaling Technology, Danvers, MA), anti-Myc tag (1:1000; Maine Medical Center Research Institute, Scarborough, ME), Ki-67 (1:150; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Standard Deviation

Journal: BMC Cancer

Article Title: Oncogenic potential of N-terminal deletion and S45Y mutant β-catenin in promoting hepatocellular carcinoma development in mice

doi: 10.1186/s12885-018-4870-z

Figure Lengend Snippet: Histological features of normal liver as well as HCCs from c-Met/β-cateninS45Y and c-Met/∆N90-β-catenin injected mice. qRT-PCR analysis of HCC tumor markers ( a ) Gross images. b H&E staining; scale bar = 200 μm. c Ki-67 IHC images; scale bar = 100 μm. d Quantification of percentage of Ki-67 (+) cells in normal liver and mouse HCC lesions. e qRT-PCR analysis of HCC tumor markers AFP and GPC3 in normal liver and mouse HCC lesions. Student’s t-test: *: P < 0.05; ***: P < 0.001; NS: Not significant

Article Snippet: After a blocking step with the 5% goat serum and Avidin-Biotin blocking kit (Vector Laboratories Inc., Burlingame, CA), the slides were incubated with the following primary antibodies: mouse anti-β-catenin (1:200; BD Transduction LaboratoriesTM, San Jose, CA), mouse anti-Glutamine Synthetase (1:500; BD Transduction LaboratoriesTM), β-catenin (D10A8) XP® Rabbit mAb (1:100; Cell Signaling Technology, Danvers, MA), anti-Myc tag (1:1000; Maine Medical Center Research Institute, Scarborough, ME), Ki-67 (1:150; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Injection, Quantitative RT-PCR, Staining

Journal: BMC Cancer

Article Title: Oncogenic potential of N-terminal deletion and S45Y mutant β-catenin in promoting hepatocellular carcinoma development in mice

doi: 10.1186/s12885-018-4870-z

Figure Lengend Snippet: Immunohistochemical patterns of β-catenin in normal liver and in HCC lesions of c-Met/β-cateninS45Y c-Met/∆N90-β-catenin mice. a and b Immunohistochemistry (IHC) of β-catenin using a mouse monoclonal antibody; scale bar = 400 μm in A; and 100 μm in B; ( c ) IHC of β-catenin using a rabbit monoclonal antibody; scale bar = 100 μm; ( d ) IHC of ectopically injected β-catenin using an anti-Myc tag antibody; scale bar = 100 μm

Article Snippet: After a blocking step with the 5% goat serum and Avidin-Biotin blocking kit (Vector Laboratories Inc., Burlingame, CA), the slides were incubated with the following primary antibodies: mouse anti-β-catenin (1:200; BD Transduction LaboratoriesTM, San Jose, CA), mouse anti-Glutamine Synthetase (1:500; BD Transduction LaboratoriesTM), β-catenin (D10A8) XP® Rabbit mAb (1:100; Cell Signaling Technology, Danvers, MA), anti-Myc tag (1:1000; Maine Medical Center Research Institute, Scarborough, ME), Ki-67 (1:150; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Immunohistochemical staining, Immunohistochemistry, Injection

Journal: BMC Cancer

Article Title: Oncogenic potential of N-terminal deletion and S45Y mutant β-catenin in promoting hepatocellular carcinoma development in mice

doi: 10.1186/s12885-018-4870-z

Figure Lengend Snippet: Biochemical analysis of HCC lesions of c-Met/β-cateninS45Y c-Met/∆N90-β-catenin mice. Western blot analysis of c-Met, p-Met, β-catenin, Myc tag, p-β-catenin (S45), p-β-catenin (S33/S37/T41), p-β-catenin (S675), p-β-catenin (S552), GS, and cyclin D1 in normal liver as well as HCC lesions from c-Met/β-cateninS45Y c-Met/∆N90-β-catenin mice. GAPDH were used as a loading control. Please note the presence of two bands for the β-catenin protein. The upper band represents endogenous β-catenin protein or β-cateninS45Y protein. The lower band represents ectopically injected ∆N90-β-catenin protein

Article Snippet: After a blocking step with the 5% goat serum and Avidin-Biotin blocking kit (Vector Laboratories Inc., Burlingame, CA), the slides were incubated with the following primary antibodies: mouse anti-β-catenin (1:200; BD Transduction LaboratoriesTM, San Jose, CA), mouse anti-Glutamine Synthetase (1:500; BD Transduction LaboratoriesTM), β-catenin (D10A8) XP® Rabbit mAb (1:100; Cell Signaling Technology, Danvers, MA), anti-Myc tag (1:1000; Maine Medical Center Research Institute, Scarborough, ME), Ki-67 (1:150; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Western Blot, Control, Injection

Journal: BMC Cancer

Article Title: Oncogenic potential of N-terminal deletion and S45Y mutant β-catenin in promoting hepatocellular carcinoma development in mice

doi: 10.1186/s12885-018-4870-z

Figure Lengend Snippet: qRT-PCR analysis of β-catenin target gene expression in c-Met/∆N90-β-catenin and c-Met/β-cateninS45Y mouse HCC samples. a qRT-PCR analysis of mRNA levels of pan-β-catenin target genes: AXIN2, LGR5, c-Myc and cyclin D1. b qRT-PCR analysis of mRNA levels of liver specific β-catenin target genes: GS, TBX3, OAT, and LECT2. Student’s t-test: * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: After a blocking step with the 5% goat serum and Avidin-Biotin blocking kit (Vector Laboratories Inc., Burlingame, CA), the slides were incubated with the following primary antibodies: mouse anti-β-catenin (1:200; BD Transduction LaboratoriesTM, San Jose, CA), mouse anti-Glutamine Synthetase (1:500; BD Transduction LaboratoriesTM), β-catenin (D10A8) XP® Rabbit mAb (1:100; Cell Signaling Technology, Danvers, MA), anti-Myc tag (1:1000; Maine Medical Center Research Institute, Scarborough, ME), Ki-67 (1:150; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Quantitative RT-PCR, Targeted Gene Expression

Journal: BMC Cancer

Article Title: Oncogenic potential of N-terminal deletion and S45Y mutant β-catenin in promoting hepatocellular carcinoma development in mice

doi: 10.1186/s12885-018-4870-z

Figure Lengend Snippet: β-cateninS45Y and ∆N90-β-catenin mutants activate TCF/LEF reporter in mouse HCC cell lines. a Western blot analysis confirmed the expression of transfected β-cateninS45Y or ∆N90-β-catenin plasmids in HCC3–4 and HCC4–4 mouse HCC cell lines. b TCF/LEF reporter activity in HCC3–4 and HCC4–4 mouse HCC cell lines following the transfection of pT3-EF1α-β-cateninS45Y and pT3-EF1α-∆N90-β-catenin, as analyzed using TopFlash/FopFlash reporter assays. Student’s t-test: *** P < 0.001

Article Snippet: After a blocking step with the 5% goat serum and Avidin-Biotin blocking kit (Vector Laboratories Inc., Burlingame, CA), the slides were incubated with the following primary antibodies: mouse anti-β-catenin (1:200; BD Transduction LaboratoriesTM, San Jose, CA), mouse anti-Glutamine Synthetase (1:500; BD Transduction LaboratoriesTM), β-catenin (D10A8) XP® Rabbit mAb (1:100; Cell Signaling Technology, Danvers, MA), anti-Myc tag (1:1000; Maine Medical Center Research Institute, Scarborough, ME), Ki-67 (1:150; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Western Blot, Expressing, Transfection, Activity Assay

Journal: BMC Cancer

Article Title: Oncogenic potential of N-terminal deletion and S45Y mutant β-catenin in promoting hepatocellular carcinoma development in mice

doi: 10.1186/s12885-018-4870-z

Figure Lengend Snippet: β-cateninS45Y and ∆N90-β-catenin mutants similarly affect proliferation and apoptosis in mouse HCC cell lines. Relative proliferation and apoptosis rates in HCC3–4 and HCC4–4 cell lines after transfection of the two β-catenin mutant forms at different time points (24 and 48 h). a proliferation; ( b ) apoptosis. Data are presented as mean ± SD. a. vs control (untransfected cells); b. vs pT3-EF1α; c. vs pT3-EF1α-β-cateninS45Y

Article Snippet: After a blocking step with the 5% goat serum and Avidin-Biotin blocking kit (Vector Laboratories Inc., Burlingame, CA), the slides were incubated with the following primary antibodies: mouse anti-β-catenin (1:200; BD Transduction LaboratoriesTM, San Jose, CA), mouse anti-Glutamine Synthetase (1:500; BD Transduction LaboratoriesTM), β-catenin (D10A8) XP® Rabbit mAb (1:100; Cell Signaling Technology, Danvers, MA), anti-Myc tag (1:1000; Maine Medical Center Research Institute, Scarborough, ME), Ki-67 (1:150; Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4 °C.

Techniques: Transfection, Mutagenesis, Control