Fig 2: CITCO treatment diminishes p38 MAPK and p65 NF-?B phosphorylation induced by TNFa in HUVEC. Cells were pretreated with vehicle (0.1% DMSO) or CITCO (3 µM) for 24h before stimulation with TNFa (20 ng/mL) for 30 min. Representative Western blots of (A) phospho p38/total p38 MAPK and (B) phospho p65/total p65 in HUVEC subjected to the different treatments. Data represent mean ± SEM of protein densitometry (n = 5 for each group). * p < 0.05 relative to vehicle group, + p < 0.05 relative to TNFa stimulated cells. (C) Representative images of NF-?B translocation into nuclei after CITCO treatment. Immunoreactivity was visualized using Alexa Fluor 488 (NF?B, green) secondary antibody. Nuclei were stained with DAPI (blue). Scale bar = 50 µm. (D) Fluorescence intensity was quantified using ImageJ. Values are expressed as mean ± SEM (n = 3 for each group). * p < 0.05 relative to vehicle group, + p < 0.05 relative to TNFa stimulated cells.

Fig 3: Inhibition of p38 MAPK signalling pathway diminishes human umbilical vein endothelial cells (HUVEC) apoptosis caused by 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD). (A) and (B) HUVECs were treated with TCDD or combined with the ERK inhibitor, PD98059 (PD, 30 µM), the JNK inhibitor, SP600125 (SP, 20 µM) or the p38MAPK inhibitor, SB203580 (SB-1, 20 µM) for 12 hrs, respectively, then stimulated with 40 nM TCDD for an additional 24 hrs. Cell viability and caspase 3 activity of HUVECs were measured. *P < 0.05 versus vehicle, # P < 0.05 versus TCDD; n = 3. (C) Effects of two different p38MAPK inhibitors (SB-1 and SB202190, SB-2) on caspase 3 activity in HUVECs treated with TCDD. *P < 0.05 versus vehicle, # P < 0.05 versus TCDD; n = 3.

Fig 4: Inhibitors of MAPK and AMPKa1/2 modulate the expression of MALT1, NDRG1, and maspin in prostate carcinoma cells. The expressions of ERK and p-ERK (A), JNK and p-JNK (C), p38 and p-p38 (E), and AMPKa1/2 and p-AMPKa1/2 (G) were determined using immunoblot assays after 20 min of 30 µM CAPE treatments with (+) or without (-) pretreatment with the indicated concentrations of inhibitors for 1 h in PC-3 cells. The quantitative data are expressed as the intensity of protein bands of the phosphorylation proteins/total protein levels relative to the control solvent-treated group. The protein levels of MALT1, NDRG1, maspin, and ß-actin in PC-3 cells after CAPE treatment with (+) or without (-) pretreatment with PD0325901 (B), SP600125 (D), SB202190 (F), or dorsomorphin (H). The quantitative data are expressed as the intensity bands of the phosphorylation proteins relative to the total protein levels or intensity of protein bands from the target genes/ß-actin relative to the control solvent-treated group. * p < 0.05, ** p < 0.01.

Fig 5: MAPK-associated signaling pathways downstream of RasGRP4 are involved in DLBCL. (a) Representative WB images showing the differences in expression and phosphorylation levels of the downstream signaling proteins between SUDHL-4 cells and normal/activated B cells. (b and c) Quantification of the levels (in a) and the phosphorylation ratio (in a). Compared with normal/activated B cells, the ratios of P-ERK1/2:ERK1/2 and P-ERK5:ERK5 were increased in DLBCL cells, while the ratio of P-JNK:JNK was decreased, and no significant change in the P-P38:P38 ratio was detected. (d) Representative WB images showing the differences in expression and phosphorylation levels of the downstream signaling proteins in RasGRP4-knockdown SUDHL-4 cells and the control group. (e and f) Quantification of the levels (in d) and the phosphorylation ratio (in d). Knockdown of RasGRP4 decreased the ERK expression and increased the JNK expression in SUDHL-4 cells; densitometry was analyzed using ImageJ software. N indicates the number of independent experiments. **P < 0.01, *P < 0.05