| Cell Line Species/Tissue: |
Rat
/
Testes |
| Transfection Reagent: |
Effectene |
Nucleic Acid:
|
DNA |
Growth Medium:
|
F-12/Dulbecco’s modified Eagle’s medium |
Percent Serum (%):
|
|
Reporter System:
|
|
Plasmid Purification Method:
|
|
Plate Format:
|
12-well plates or cover glass |
Number of Cells:
|
|
Percent Confluence(%):
|
6x10e5 |
Amount of Nucleic Acid (µg):
|
~0.3µg of plasmid DNA |
Amount of Enhancer (µl):
|
|
Amount of Reagent (µl):
|
2.4 µl |
Complex Incubation on Cells (hrs):
|
24 h |
Analysis Performed Post-Transfection (hrs):
|
|
Transfection Efficiency (%):
|
~15% |
| |
| Any modifications to the protocol?:
|
| |
| Notes: |
| Primary Sertoli cells cultured in vitro were transfected with plasmids containing the full-length TGF-ß3 (pCIneo/TGF-ß3) with ~15% efficiency. About 24 h after transfection, both the steady-state mRNA and protein levels of TGF-ß3 increased significantly. |
| References: |
| Differential Interactions between Transforming Growth Factor-ß3/TR1, TAB1, and CD2AP Disrupt Blood-Testis
Barrier and Sertoli-Germ Cell Adhesion
Weiliang Xia, Dolores D. Mruk, Will M. Lee, and C. Yan Cheng
THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 281, NO. 24, pp. 16799–16813, June 16, 2006
|