FIG 8.

PLSCR1 represses transcription from the CRE- and HCMV MIEP-regulated reporter constructs. (A) HEK-293 cells (1 × 10⁵) were plated in 24-well cell culture plates and transfected with 40 ng of CRE-responsive reporter plasmid (pCRE-Luc), 40 ng of TAL-pGL4.70 (53), and 300 ng of pcDNA3 or S-PLSCR1 using jetPEI. After 24 h of transfection, the cells were treated with or without 5 μM Forskolin (Cayman Chemical) for 16 h. Then the cells were lysed, and their luciferase activity levels were determined. The firefly luciferase/Renilla luciferase activity ratio of cells transfected with the vector only was defined as 100%. Data represent the average relative values from triplicates, and error bars indicate standard deviations. **, P < 0.01 by Student's t test. (B) HeLa-PLSKO cells (5 × 10⁴) were plated in 24-well cell culture plates and transfected with 2 ng of pMIEP-Luc, 40 ng of pGL4.74 (Promega), and 350 ng of pcDNA3 or S-PLSCR1 using TransIT-X2. After 24 h of transfection, the cells were treated with or without 5 μM Forskolin for 16 h. Then the cells were lysed, and their luciferase activity levels were determined. The firefly luciferase/Renilla luciferase activity ratio of cells transfected with the vector only was defined as 100%. Data represent average relative values from triplicates, and error bars indicate standard deviations. **, P < 0.01 by Student's t test.