epithelial cells human telomerase reverse transcriptase htert  (ATCC)

Journal: Molecular Medicine Reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: Immortalised human epithelial cells (human telomerase reverse transcriptase-human pancreatic nestin-expressing ductal cells) were cultured in 25 cm 2 flasks and supplemented with recombinant TF (0.5 U/ml) or were untreated. The cells were harvested on the indicated weeks, total RNA was isolated from one group and the mRNA quantified by RT-qPCR, against β-actin. Other aliquots were analysed by western blotting. The data show the amounts of (A) Inhibitor of CDK p16 INKa mRNA (n=3), and (B) the relative amounts of p16 INKa protein (n=4) calculated from (C) the western blots of p16 INKa protein (using a goat anti-human antibody) and against GAPDH. TF, tissue factor; Wk, week.

Article Snippet: The immortalised pancreatic epithelial cells [human telomerase reverse transcriptase (hTERT)-human pancreatic nestin-expressing ductal cells (HPNE)] were purchased from American Type Culture Collection (ATCC).

Techniques: Reverse Transcription, Expressing, Cell Culture, Recombinant, Isolation, Quantitative RT-PCR, Western Blot

Journal: Molecular Medicine Reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: Immortalised human epithelial cells (human telomerase reverse transcriptase-human pancreatic nestin-expressing ductal cells) were cultured in 25 cm 2 flasks and supplemented with recombinant TF (0.5 U/ml) or were untreated. The cells were harvested on the indicated wks, total RNA was isolated from one group and the mRNA quantified by RT-qPCR, against β-actin. Other aliquots were analysed by western blotting. The data show the amounts of (A) CDK interacting protein/Wildtype p53-activated fragment p21 CIP1/WAF1 mRNA (n=3), and (B) the relative amounts of p21 CIP1/WAF1 protein (n=3) calculated from (C) the western blots of p21 CIP1/WAF1 protein (using a mouse anti-human antibody) and against GAPDH. TF, tissue factor; Wk, week.

Article Snippet: The immortalised pancreatic epithelial cells [human telomerase reverse transcriptase (hTERT)-human pancreatic nestin-expressing ductal cells (HPNE)] were purchased from American Type Culture Collection (ATCC).

Techniques: Reverse Transcription, Expressing, Cell Culture, Recombinant, Isolation, Quantitative RT-PCR, Western Blot

Journal: Molecular Medicine Reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: Immortalised human epithelial cells (human telomerase reverse transcriptase-human pancreatic nestin-expressing ductal cells) were cultured in 25 cm 2 flasks and supplemented with recombinant TF (0.5 U/ml) or were untreated. The cells were harvested on the indicated weeks, total RNA was isolated from one set and the mRNA quantified by RT-qPCR, against β-actin. Other aliquots were analysed by western blotting. The data show the amounts of (A) cyclin E mRNA (n=3), and (B) the relative amounts of Cyclin E protein (n=3) calculated from (C) the western blots of Cyclin E protein (using a rabbit anti-human antibody) and against GAPDH. TF, tissue factor.

Article Snippet: The immortalised pancreatic epithelial cells [human telomerase reverse transcriptase (hTERT)-human pancreatic nestin-expressing ductal cells (HPNE)] were purchased from American Type Culture Collection (ATCC).

Techniques: Reverse Transcription, Expressing, Cell Culture, Recombinant, Isolation, Quantitative RT-PCR, Western Blot

Journal: Molecular Medicine Reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: Immortalised human epithelial cells (human telomerase reverse transcriptase-human pancreatic nestin-expressing ductal cells) were supplemented with recombinant TF (0.5 U/ml) or were untreated. The cells were harvested on week 4, gDNA was extracted from the cells (3×10 4 cells) and bisulphite conversion of the gDNA (750 ng) was carried out using the MethylDetector Bisulfite Modification Kit. DNA was also extracted from HT-29 cells (methylated control) and HDBEC (unmethylated control) and processed as aforementioned. The modified DNA (10 ng/reaction) was amplified with methylation-specific, and unmethylated-specific sets primers to the p16 gene promoter region. Each of the nested amplification steps was carried out for 35 cycles at an annealing temperature of 60°C. Aliquots (4 µl) from the outer reactions were then used as the template for the inner PCR reactions using primers specific for methylated and unmethylated DNA. Both amplicons generated bands of 149 bp. A control β-actin sample was also amplified and examined alongside. (A) The products were examined by 2% (w/v) agarose gel electrophoresis and (B) the band intensities determined and the ratios of the methylated:unmethylated DNA calculated (n=3). TF, tissue factor; gDNA, genomic DNA.

Article Snippet: The immortalised pancreatic epithelial cells [human telomerase reverse transcriptase (hTERT)-human pancreatic nestin-expressing ductal cells (HPNE)] were purchased from American Type Culture Collection (ATCC).

Techniques: Reverse Transcription, Expressing, Recombinant, Modification, Methylation, Control, Amplification, Generated, Agarose Gel Electrophoresis

Journal: Molecular Medicine Reports

Article Title: Tissue factor signalling modifies the expression and regulation of G1/S checkpoint regulators: Implications during injury and prolonged inflammation

doi: 10.3892/mmr.2024.13404

Figure Lengend Snippet: Immortalised human epithelial cells (human telomerase reverse transcriptase-human pancreatic nestin-expressing ductal cells) were cultured in 25 cm 2 flasks and supplemented with recombinant TF (0.5 U/ml) or were untreated. The cells were harvested on the indicated wks, total RNA was isolated from one set and the mRNA quantified by RT-qPCR, against β-actin. Other aliquots were analysed by western blotting. The data show the amounts of (A) Alternative reading frame p14 ARF mRNA (n=3), and (B) the relative amounts of p14 ARF protein (n=3) calculated from (C) the western blots of p14 ARF protein (using a rabbit anti-human antibody) and against GAPDH. TF, tissue factor; Wk, week.

Article Snippet: The immortalised pancreatic epithelial cells [human telomerase reverse transcriptase (hTERT)-human pancreatic nestin-expressing ductal cells (HPNE)] were purchased from American Type Culture Collection (ATCC).

Techniques: Reverse Transcription, Expressing, Cell Culture, Recombinant, Isolation, Quantitative RT-PCR, Western Blot