Journal: Thrombosis research

Article Title: Enhanced functional recombinant factor VII production by HEK 293 cells stably transfected with VKORC1 where the gamma-carboxylase inhibitor calumenin is stably suppressed by shRNA transfection.

doi: 10.1016/j.thromres.2007.11.002

Figure Lengend Snippet: Figure 1 Stable silencing of calumenin in HEK 293 cells with HuSH 29mer shRNA constructs against human calumenin. Four pRS shRNA vectors with different 29mer oligonucleo- tides called CALU1, 2, 3 and 4 were used individually to stably transfect HEK 293 cells stably overexpressing r-hFVII and VKORC1 (see Materials and methods). The cellular concen- tration of calumenin in the various CALU transfected cells was measured after Western blotting and compared to calumenin concentration in cells transfected with the vector (control, 100%). Aliquots from the media harvested from the different cell cultures were loaded in each lane. Affinity purified antibodies against the 47 kDa calumenin were used to identify the protein. The same blot was stripped and reprobed with a β-Actin antibody to correct for equal protein loading in each lane. Quantification of the various immunor- eactive bands was made by chemiluminescence (see Materi- als and methods).

Article Snippet: Packaging of human calumenin shRNA into replication deficient retrovirus The kit HuSH 29mer shRNA Construct against CALU (TR314215) against human calumenin was purchased from ORIGEN (Rockville, MD).

Techniques: shRNA, Construct, Stable Transfection, Transfection, Western Blot, Concentration Assay, Plasmid Preparation, Control, Affinity Purification

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Caveolae as a target to quench autoinduction of the metastatic phenotype in lung cancer

doi: 10.1007/s00432-015-2074-3

Figure Lengend Snippet: The response to the same treatments in CaLu-1 cells where significant changes to gene expression can be seen. Zoledronic acid displays a modest effect in FOXC2 (transcription factor FKH-14 associated with EMT) and FN1 (a fibronectin)

Article Snippet: MDA-MB-231 cells were obtained from Cell Lines Service, Eppenheim, Germany, and CaLu-1 cells were obtained from ECCAC, UK.

Techniques: Gene Expression

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Caveolae as a target to quench autoinduction of the metastatic phenotype in lung cancer

doi: 10.1007/s00432-015-2074-3

Figure Lengend Snippet: The effects of the treatments on CaLu-1 cells. The data are filtered for significance ( p < 0.05) and by the subset of genes associated with signalling. X-axis shows log2 fold change. Transcription of FYN and ABL1 is reduced; both FYN and ABL proteins mediate transduction at caveolae. PDGF and PDGF/R are likewise down-regulated. The down-regulated genes at the top of the graph are associated with caveolae, while the up-regulated genes at the lower part of the graph are associated with rafts

Article Snippet: MDA-MB-231 cells were obtained from Cell Lines Service, Eppenheim, Germany, and CaLu-1 cells were obtained from ECCAC, UK.

Techniques: Transduction

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Caveolae as a target to quench autoinduction of the metastatic phenotype in lung cancer

doi: 10.1007/s00432-015-2074-3

Figure Lengend Snippet: CAV1 down-regulation is mirrored in the CAV-1 protein assay only in CaLu-1 cells. Expressed protein levels in MDA-MB-231 cells are not significantly different from the control groups. Error bars are ±SE

Article Snippet: MDA-MB-231 cells were obtained from Cell Lines Service, Eppenheim, Germany, and CaLu-1 cells were obtained from ECCAC, UK.

Techniques: Control

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Caveolae as a target to quench autoinduction of the metastatic phenotype in lung cancer

doi: 10.1007/s00432-015-2074-3

Figure Lengend Snippet: MDA-MB-231 cells share a reduction in half of the signal proteins compared to CaLu-1. In only the lung cells the receptors for EGF, PDGF, androgens and VEGF are affected by the treatment. Error bars are ±SE

Article Snippet: MDA-MB-231 cells were obtained from Cell Lines Service, Eppenheim, Germany, and CaLu-1 cells were obtained from ECCAC, UK.

Techniques:

Journal: Thrombosis research

Article Title: Enhanced functional recombinant factor VII production by HEK 293 cells stably transfected with VKORC1 where the gamma-carboxylase inhibitor calumenin is stably suppressed by shRNA transfection.

doi: 10.1016/j.thromres.2007.11.002

Figure Lengend Snippet: Figure 1 Stable silencing of calumenin in HEK 293 cells with HuSH 29mer shRNA constructs against human calumenin. Four pRS shRNA vectors with different 29mer oligonucleo- tides called CALU1, 2, 3 and 4 were used individually to stably transfect HEK 293 cells stably overexpressing r-hFVII and VKORC1 (see Materials and methods). The cellular concen- tration of calumenin in the various CALU transfected cells was measured after Western blotting and compared to calumenin concentration in cells transfected with the vector (control, 100%). Aliquots from the media harvested from the different cell cultures were loaded in each lane. Affinity purified antibodies against the 47 kDa calumenin were used to identify the protein. The same blot was stripped and reprobed with a β-Actin antibody to correct for equal protein loading in each lane. Quantification of the various immunor- eactive bands was made by chemiluminescence (see Materi- als and methods).

Article Snippet: Packaging of human calumenin shRNA into replication deficient retrovirus The kit HuSH 29mer shRNA Construct against CALU (TR314215) against human calumenin was purchased from ORIGEN (Rockville, MD).

Techniques: shRNA, Construct, Stable Transfection, Transfection, Western Blot, Concentration Assay, Plasmid Preparation, Control, Affinity Purification