FIGURE 6.

Artesunate (AS) inhibits the physical interaction between VDR and NF‐κB p65 in LPS‐tolerant macrophages. RAW264.7 cells were treated as described in the legend of Figure 2d. (a) The cytoplasm (a1) and nuclear (a2) lysate were used for an IP experiment using anti‐VDR antibodies and the associated NF‐κB p65 (p65) was detected by immunoblotting (IB). (b) Immunostaining to observe the co‐localization of p65 and VDR. p65 was probed using Alexa Fluor 488 (green). VDR was probed using Alexa Fluor 555 (red). Representative images are shown (bar = 5 μm) (b1). The co‐localization of VDR and p65 (b2) and the karyoplasmic ratio of p65 (b3) was quantified from 100 cells (normalized to medium). (c) The p65 level in the nuclear lysate was detected using elisa and WB. (d) Change in the p65 level in Vdr‐KD (d1) or Vdr‐OE (d2) LPS‐tolerant RAW264.7 cells treated with AS. (e) Change in the TNF‐α level in p65‐KD (e1) or p65‐OE (e2) LPS‐tolerant RAW264.7 cells treated with AS (n = 5). One‐way ANOVA followed by Tukey's post hoc test; ns, not significant; ^* P < 0.05