Effect of a 6-h incubation with Sch A and Sch B in lipopolysaccharide (LPS)-activated pro-inflammatory signals in RAW264.7 macrophages.

<p>After incubation with Sch A and Sch B for 6 h, RAW264.7 macrophages were challenged with LPS (1 μg/mL) for 15 min. (A, B) The levels of phospho-JNK1/2 (and phospho-p38) and total-JNK1/2 (and total p38) were measured using Western blot analysis. The ratio of phoshos-JNK1/2 to total-JNK1/2 as well as the ratio of phospho-p38 to total p38 (which are indicative of JNK1/2 and p38 activation) were estimated and expressed as % control, by normalizing with the value of the non-LPS group (JNK1/2: left panel, p38: right; upper panel). (C) After transfection with NF-κB-luciferase reporter in RAW264.7 macrophages, the RAW264.7 cells were incubated with Sch A or Sch B, as described in Materials and Methods. NF-κB-luciferase reporter, which consists of an NF-κB- responsive firefly luciferase construct and a constitutively expressed Renilla luciferase. Following exposure to LPS for 1 h, luciferase activities in the cell lysates were measured. Renilla luciferase expression was used to normalize the varying transfection efficiencies among the samples. NF-κB reporter activity was expressed as % control, by normalizing with the value of the non-LPS control. Value given are means ± SEM, with n = 3. * Significantly different from the non-LPS control; # significantly different from the LPS control.