(A) Heat map (reverse phase protein array) of protein expression and phosphorylation level in RAW 264.7 cells stimulated with vehicle (PBS), nCB (105 ng/ml), and PEG-CB (105 ng/ml). p: phosphorylated. Blue is relatively low (−0.5) and yellow high (0.5) based on log2 ratio of the value for expression level. (B) RAW 264.7 cells under indicated conditions immunostained for nuclear DNA (DAPI, blue) and anti-γH2AX (green) to detect double strand break (DSB). Scale bar: 50 μm. (C) Quantitative summary of panel B indicating the percentage γH2AX positive RAW cells in indicated groups. (D) IL-6 concentration detected by ELISA after 48 hr in the supernatant of MDDC treated with CB or LPS in the presence of increasing dose of Nu7026 or vehicle (DMSO). (E) IL-17A concentration detected by ELISA after 72 hr co-culture of splenic CD4 T cells and lung CD11c+ cell isolated from the mice after challenged with PBS or nCB and anti-CD3 (1 μg/ml) in the presence of Nu7026 (100 nM), Ku55933 (100 nM), or vehicle control (DMSO). (F) Western blot of protein extracted from BMDC treated with different concentration of nCB targeting phosphorylated-Erk. Data are representative of two independent experiments. (G) IL-6 concentration detected by ELISA in the supernatant of MDDC treated with nCB in the presence of increasing dose of U0126 (MEK1/2 inhibitor) for 48 hr. n = 4 to 7 per group and data are mean ± SEM and representative of two independent experiments (C, D, E, G). ***p < 0.001, **p < 0.01 as determined by the one-way ANOVA and Bonferroni's multiple comparison test.