Fig 2: Nuclear levels of p50, p38 and phosphorylated p38 in the nuclear fraction of N9 cells exposed for 6 h to Hg2+ (5 μM) or Lipopolysaccharide (LPS; 300 ng/mL). (a) Western blot results for p50 (50 kDa), p38 (38 kDa) and phosphorylated p38 (p-p38; 38 kDa) using Lamin B (67 kDa) as a nuclear marker. (b) Band intensity analysis for p38 and p-p38. Data represent the mean ± S.E. of 3 independent experiments. * significantly different (p < 0.05) from non-treated control (Ct); ** very significantly different (p < 0.01) from non-treated control (Ct).

Fig 3: Effects of AM251 on the inflammatory response and MAPK‐related factor levels in the liver tissues of db/db mice. A, Hepatic mRNA levels of TNF‐α, IL‐6, MCP‐1, and IFNγ were measured by quantitative real‐time polymerase chain reaction. B, Hepatic protein phosphorylation levels of p38, ERK, and JNK were detected using Western blot analysis. *P < .05 vs Lean control mice (n = 7); † P < .05 vs db/db mice (n = 8). ERK, extracellular signal‐regulated kinase; IFNγ, interferon γ; IL‐6, interleukin‐6; JNK, c‐Jun N‐terminal kinase; MAPK, mitogen‐activated protein kinase; MCP‐1, monocyte chemoattractant protein‐1; mRNA, messenger RNA; MW, molecular weight; TNF‐α, tumor necrosis factor‐α

Fig 4: Transcript (mRNA) levels for IL1-ß (a), TNF-alpha (b) and iNOS (c) following exposure of N9 cells for 24 h to Hg2+ (5 μM) alone or after pre-exposure to either the p38 inhibitor SB 239063 (4 h, 100 μM), N-acetylcysteine (NAC; 2.5 mM, 24 h) or sodium selenite (Se; 0.1 μM, 24 h). Cells exposed to Lipopolysaccharide (LPS) were used as a positive control. Data represent the mean ± S.E. of 3–5 independent experiments. * significantly different (p < 0.05) from non-treated control (Ct); ** very significantly different (p < 0.01) from Ct; # significantly different (p < 0.05) from cells treated with Hg2+; ## very significantly different (p < 0.01) from cells treated with Hg2+.

Fig 5: NED416 activates mitogen-activated protein kinase signaling proteins. (A) Expression of p-ERK, p-JNK and p-p38 protein kinases after NED416 treatment in mouse skin samples. Quantification of the activation of (B) ERK, (C) JNK and (D) and p38 upon NED416 treatment. Data are presented as the mean ± SEM. *P<0.05, **P<0.01, ***P<0.001 vs. 0%. NED416, (E)-3-(2,4-dichlorophenyl)-N-phenylacrylamide; p, phosphorylated.