Fig 2: OJS treatment inhibits the activation of MAPK and NF-?B pathways in cisplatin-induced AKI. (A) The phosphorylation levels of p38, ERK1/2, and JNK were analyzed by western blotting. Total p38, ERK1/2, and JNK were used as loading controls. (B) The relative density ratio of phosphorylated form/total form. (C) The degradation of I?-Ba was analyzed by western blotting. ß-actin was used as a loading control. (D) The relative density ratio of I?-Ba/ß-actin. Data are represented as mean ± S.E.M. (n = 6; * indicates p < 0.05 vs. saline-treated control group, † indicates p < 0.05 vs. cisplatin treatment alone).

Fig 3: The EP3-dependent p38MAPK/Bcl-2 pathway is activated in 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD)-treated human umbilical vein endothelial cells (HUVECs). (A) The mRNA levels of Bcl-2 in TCDD-treated HUVECs were determined by qRT-PCR. TCDD dose is shown in the X-axis. *P < 0.05 versus the 0 nM group; n = 3. (B) Protein levels of Bcl-2 in HUVECs treated with or without 40 nM TCDD for 24 hrs were analysed by Western blot. (C–E) HUVECs were transfected with pcDNA3.1-Bcl-2 (pc-Bcl2) plasmid or vector and then incubated with TCDD (40 nM) or DMSO for 24 hrs. Cells were harvested for Bcl2 protein expression analysis (C), cell viability (D) and caspase 3 activity assays (E). *P < 0.05 versus vector + vehicle, # P < 0.05 versus vector + TCDD; n = 3. (F–G) HUVECs were pre-treated with the ERK inhibitor, SB203580 (F) or L798106 (G) for 12 hrs, respectively, and then stimulated with 40 nM TCDD for an additional 24 hrs. Protein levels of phosphorylated p38 MAPK, total p38 MAPK and Bcl-2 were examined by Western blot.

Fig 4: Attenuated monocyte recruitment molecule expression in p38a deficient endothelial cells, in vitro.(A) Flow cytometric analysis of primary lung endothelial cells to check for purity of population, after treatment with HTNC for 16 hrs. Cells were stained with CD146 FITC, specific for MLECs. Leukocytes isolated from blood were used as negative controls for CD146 staining (data not shown). (B) Immunoblotting for p38a and total p-p38 on MLEC protein extracts. (C) Relative mRNA expression levels of adhesion molecule VCAM1 and chemokines IP-10, MCP-1 and Gro-KC in MLECs. Cells were isolated from the lung of p38aFL/FL/ApoE-/- and ApoE-/- mice, passaged twice and treated with HTNC for 16 hrs to induce cre recombination. Data shown are representative of two separate experiments. Error bars represent SD.

Fig 5: CAPE increases phosphorylation of ERK, p38, JNK, and AMPKa1/2 in PC-3 cells. (A) Time courses of ERK, p-ERK, p38, p-p38, JNK, p-JNK, AMPKa1/2, and p-AMPKa1/2 expressions were determined using immunoblot assays after various periods of 30 µM CAPE treatments in PC-3 cells. (B) Dose responses of ERK, p-ERK, p38, p-p38, JNK, p-JNK, AMPKa1/2, and p-AMPKa1/2 were determined using immunoblot assays after 30 min of various concentrations of CAPE treatments in PC-3 cells. The quantitative data are expressed as the intensity bands of the phosphorylation proteins relative to the total protein levels for ERK (C,D), p38 (E,F), JNK (G,H), and AMPK (I,J). * p < 0.05, ** p < 0.01.