Fig. 2
Identification of standard screening method based on RAW264.7 Cell line stably expressing NF-κB-dependent SEAP reporter Gene. a Standard curve of alkaline phosphatase, y = 0.0315x, R2 = 0.9947. b Relationship between OD405 and cell number/well of Raw264.7 cells stably expressing NF-κB dependent SEAP reporter gene (RAW264.7-pNF-κB-SEAP cells)treated with or without LPS. R2 = 0.9395 (the cells were treated with LPS), and R2 = 0.9910 (the cells were treated without LPS). c The effect of RAW264.7-pNF-κB-SEAP cells were treated with different concentrations of LPS for 24 h menifested as column chart of relative activity of SEAP and dose-response stimulation curve with common logarithm of LPS concentration as the ordinate and the stimulation ratio (%) as the abscissa, R2 = 0.9643. d The RAW264.7-pNF-κB-SEAP cells were treated with PDTC at concentrations of 0.1, 1, 10, and 100 μM or 1‰ DMSO (v/v) (as control) for 6 h and then treated with LPS in 100 ng/mL for 24 h. The effect of PDTC on RAW264.7-pNF-κB-SEAP cells was manifested as column chart of relative activity of SEAP and the dose-response inhibition curve between the log10 concentrations of PDTC and the inhibition ratio calculated from SEAP relative activity, R2 = 0.9802, their relation according to the common logarithmic equation. e The RAW264.7-pNF-κB-SEAP cells were treated with BAY117085 at concentrations of 0.1, 1, 10, and 50 μM or 1‰ DMSO (v/v) (as control) for 6 h and then treated with LPS in 100 ng/mL for 24 h. The effect of BAY117085 on RAW264.7-pNF-κB-SEAP cells was manifested as column chart of relative activity of SEAP and the dose-response inhibition curve between the log10 concentrations of BAY117085 and the inhibition ratio calculated from SEAP relative activity, R2 = 0.9735, their relation according to the common logarithmic equation. The “*” indicates that the column had significant difference compared with the control, the others marked as “#” and “^” indicate that the column showed significant difference compared with the column on the left