Stable ΔN-IκBα Expressing HOS-G (HOS-G/ΔN-IκBα) Cell Line
|T6108||1x106 cells / 1.0 ml|
|Description||Human Osetosarcoma (HOS) cells were transfected with a Lentivirus vector to stably express a degradation resistant I-κBα (?N-I-κBα) under the control of a SV40 promoter. Successfully transfected cells were selected for puromycin resistance. As a result of the constitutive repression of NF-κB by the degradation resistant inhibitor ?N-I-κBα, these cells are resistant to senescence caused by infection with human T-lymphotropic virus type 1 (HTLV-1). HOS-G/ΔN-IκBα line can be productively infected by HTLV-1 and can spread HTLV-1 to susceptible cells. This line also contains a reporter cassette consisting of 18 copies of Tax-inducible HTLV-1 21bp repeat, the HTLV-1 TATA element, the complete R region, and part of the U5 sequence fused with enhanced green fluorescence protein (eGFP). As a result of this reporter element the expression of eGFP is a positive indicator of successful HTLV-1 infection in cells. It is suggested that this cell line can be used to study of HTLV-1 replication in cell culture.|
|Species||Human (H. sapiens)|
|Seeding Density||Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.|
For Research Use Only
|Unit quantity||1x106 cells / 1.0 ml|
|Cell Type||Drug Discovery Cells|
δN-I-κBα , eGFP
|Propagation Requirements||The base medium for this cell line is Prigrow III available at abm, Cat. No. TM003 . To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999) to final concentration of 10%, 2mM L-glutamine (G275), Penicillin/Streptomycin (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: 37.0|
1) Verified the expression of GFP upon HTLV-1 infection via imaging (Figure 1). 2) Verified the expression of δN-IκBα in Western blot (Figure 1). 3) Verified that cell line continues to propagate after HTLV-1 infection (Figure 2). 4) Confirmed the expression of viral genes Tax, Rex and Gag through Western Blot analysis ( HTLV-1 proteins) (Figure 3A). 5) Verified viral spread to B-cells through Electron microscopy (Figure 3B). 6) Showed cells could spread from one HOS line to another as well as to T-cell reporter line by imaging RFP expression and measuring Luciferase activity of the T-cell reporter line (Figure 5).
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3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.
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|Depositor||The Henry M. Jackson Foundation for the Advancement of Military Medicine|
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