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. 2017 Nov 15;7(12):1972–1981. doi: 10.1002/2211-5463.12340

Figure 2.

Figure 2

The effects of EGCG3″Me on PGE 2 production and the mRNA expression of related genes in osteoblasts. (A) The levels of PGE 2 in cultured medium of osteoblasts. POBs were cultured for 24 h with or without LPS (1 ng·mL−1) and EGCG3″Me (30 μm), and the conditioned medium was collected to measure the levels of PGE 2. (B) The mRNA expression of COX‐2 (Cox2), mPGES‐1 (mPges1), and RANKL (Rankl) in osteoblasts was analyzed using real‐time PCR. POBs were cultured for 24 h with or without LPS (1 ng·mL−1) and EGCG3″Me (30 μm), and total RNA was isolated for real‐time PCR. (C) IKK activity was determined in vitro with or without EGCG3″Me (1 mm) by the IKK activity assay kit using IKKβ, IκBα, and anti‐phospho‐IκBα antibody. IKK activity was expressed as the % of the control without EGCG3″Me. (D) NF‐κB‐mediated transcriptional activity was measured with or without EGCG3″Me (30 μm). Plasmid pNFkB‐TA‐Luc (0.4 μg) and the pGL4.74[hLuc/TK] plasmid (40 ng) were transfected into mouse POBs, and the POBs were cultured for 24 h with or without LPS (1 ng·mL−1) and EGCG3″Me (30 μm). The luciferase activity was measured with the Dual‐luciferase Reporter Assay system. A significant difference between the two groups was indicated; **P < 0.01 and ***P < 0.001 vs. control, ## P < 0.01 and ### P < 0.001 vs. LPS.