728196.fig.002a
(a)
728196.fig.002b
(b)
728196.fig.002c
(c)
Figure 2: Inhibition of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression by Zuonin B. RAW264.7 cells were pretreated with different concentrations of Zuonin B for 1 h and stimulated with LPS (1 μg/mL) for a further 24 h. Equal amounts of protein in cell lysates were electrophoresed, and the protein expression levels of iNOS (a) and COX-2 (b) determined using specific antibodies for iNOS and COX-2. The respective levels of β-actin were used to confirm equal amounts of protein loading for electrophoresis. Interleukin COX-2 and iNOS mRNA levels in lung tissue, (c) Cells were cultured for 24 h with LPS (1 μg/mL), fixed, permeabilized, and incubated with rabbit polyclonal anti-iNOS and COX-2 antibody, followed by Texas-red conjugated anti-rabbit Ig (red). The nuclei of the corresponding cells were visualized with 4′,6-diamidino-2-phenylindole (DAPI) (magnification 400). NC: untreated control cells; LPS: LPS only treatment.