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. Author manuscript; available in PMC: 2008 Aug 15.
Published in final edited form as: Mol Cell Endocrinol. 2007 May 29;274(1-2):35–42. doi: 10.1016/j.mce.2007.05.013

Fig.2. Effect of LPS on GHR promoter is not secondary to release of cytokines into the cell culture medium.

Fig.2

(A) HEK 293T cells were transfected with L2-promoter-luciferase reporter (pGL3-L2-2.0 kb), pRL-TK (internal control), wild type TLR-4 (0.1 μg/well), and wild type MD2 (0.1 μg/well) plasmids. 48 h after transfection, cells were stimulated with LPS (1 μg/ml) for 5-6 h. After stimulation, the cells were harvested and the supernatant analyzed for cytokine levels. Results are expressed as mean ±SEM; n= 3. (B) HEK 293T cells were transfected with a L2-promoter-luciferase reporter (pGL3-L2-2.0 kb), pRL-TK (internal control), wild type TLR-4 (0.1 ng/well), and wild type MD2 (0.1 ng/well). 48 h after transfection, cells were stimulated for 5-6 h with TNF-α (10 or 40 pg/ml), LPS (1 μg/ml), or a combination of LPS (1 μg/ml) and TNF-α (10 ng/ml). After stimulation, the cells were harvested and the luciferase activity measured. Results are expressed as mean ±SEM; n= 3. p < 0.05 (Mann-Whitney U test) compared to the luciferase activity in the absence of LPS (vehicle).