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. 2013 Dec 10;171(1):237–252. doi: 10.1111/bph.12440

Figure 2.

Figure 2

Andrographolide induces HO-1 expression. (A–C) Induction of HO-1 promoter activity (A), HO-1 mRNA transcription (B) and HO-1 protein synthesis (C) by andrographolide in Ava5 cells. The HO-1 promoter-linked firefly luciferase (FLuc) vector, pHO-1-Luc, was transiently transfected into Ava5 cells. Each transfection mixture contained 0.1 μg pSEAP reporter plasmid as a control of transfection efficiency. The transfected cells were then treated with andrographolide at different concentrations (0–10 μM) for 3 days. The HO-1 promoter activity was assayed for luciferase activity and presented as fold activation relative to a parental Huh-7 cell control (defined as 1) following normalization against SEAP activity. The HO-1 RNA level was normalized against the cellular gapdh mRNA level and presented as the percentage change relative to parental Huh-7 cells (defined as 100%). Protein synthesis was detected by Western blotting with anti-HO-1 antibody. Equal loading of cell lysates was confirmed by probing the same blot with anti-GAPDH antibody. The experiments were performed in triplicate, and error bars indicate mean ± SD. *P < 0.05, **P < 0.01.