Fig. 4
Intra-cellular signaling pathways in HUVECs stimulated with SSc-ICs, PAPS-ICs, SLE-ICs, or NHS-ICs. Endothelial cells were incubated with SSc-ICs, PAPS-ICs, SLE-ICs, or NHS-ICs (1:2 dilution). IL-1β (50 U/ml) was used as a positive control. Results are expressed as the ratio of phosphorylated to non-phosphorylated forms, evaluated using the ImageJ software. Western Blotting images are representative of a single experiment. pNFκB: phosphorylated NFκB; p38MAPK: phosphorylated p38MAPK; pp46SAPK-JNK: phosphorylated p46SAPK-JNK; pp54SAPK-JNK: phosphorylated p54SAPK-JNK; pAkt: phosphorylated Akt. Histograms represent mean + standard error of the mean (SEM). *p < 0.01; **p < 0.001; ***p < 0.0001 versus medium. a pNFκB/NFkB. IL-1β [7.75 ± 0.49] versus medium [1.02 ± 0.05] (p < 0.0001) versus NHS-ICs (p < 0.0001). ATA-ICs [5.00 ± 0.55] versus medium (p < 0.0001) and versus NHS-ICs [22.75 ± 7.59] (p < 0.0001). ACA-ICs [3.00 ± 1.39] versus medium (p < 0.01) versus NHS-ICs (p = N.S.). ARA-ICs [2.17 ± 1.55] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). Anti-Th/To-ICs [2.98 ± 1.75] versus medium (p < 0.01) versus NHS-ICs (p = N.S.). PAPS-ICs [2.13 ± 0.26] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). SLE-ICs [1.65 ± 0.13] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). NHS-ICs [1.25 ± 0.21] versus medium (p = N.S.). b pp38/p38. IL-1β [31.98 ± 9.37] versus medium [1.4 ± 0.4] (p < 0.0001) versus NHS-ICs (p < 0.0001). ATA-ICs [26.75 ± 7.3] versus medium (p < 0.0001) and versus NHS-ICs [22.75 ± 7.59] (p < 0.001). ACA-ICs [22.43 ± 9.57] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). ARA-ICs [26.15 ± 5.27] versus medium (p < 0.001) versus NHS-ICs (p < 0.001). Anti-Th/To-ICs [24.65 ± 12.77] versus medium (p < 0.001) versus NHS-ICs (p < 0.001). PAPS-ICs [21.93 ± 7.77] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). SLE-ICs [11.63 ± 9.14] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). NHS-ICs [3.2 ± 0.85] versus medium (p = N.S.). c pp46SAPK-JNK/p46SAPK-JNK. IL-1β [1.98 ± 0.17] versus medium [1.03 ± 0.05] (p < 0.0001) versus NHS-ICs (p < 0.0001). ATA-ICs [1.85 ± 0.19] versus medium (p < 0.0001) and versus NHS-ICs [22.75 ± 7.59] (p < 0.0001). ACA-ICs [1.65 ± 0.13] versus medium (p < 0.001) versus NHS-ICs (p < 0.0001). ARA-ICs [1.60 ± 0.48] versus medium (p < 0.01) versus NHS-ICs (p < 0.001). Anti-Th/To-ICs [1.23 ± 0.22] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). PAPS-ICs [1.22 ± 0.09] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). SLE-ICs [1.13 ± 0.19] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). NHS-ICs [0.88 ± 0.32] versus medium (p = N.S.). d pp54SAPK-JNK/p54SAPK-JNK. IL-1β [20.88 ± 1.56] versus medium [1.05 ± 0.06] (p < 0.0001) versus NHS-ICs (p < 0.0001). ATA-ICs [7.30 ± 0.63] versus medium [21.75 ± 6.24] (p < 0.0001) and versus NHS-ICs [22.75 ± 7.59] (p < 0.0001). ACA-ICs [3.75 ± 0.59] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). ARA-ICs [2.52 ± 0.41] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). Anti-Th/To-ICs [2.02 ± 0.60] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). PAPS-ICs [3.98 ± 0.24] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). SLE-ICs [3.05 ± 2.17] versus medium (p < 0.01) versus NHS-ICs (p = N.S.). NHS-ICs [1.73 ± 0.28] versus medium (p = N.S.). e pAkt/Akt. IL-1β [1.90 ± 0.39] versus medium [1.02 ± 0.05] (p < 0.01) versus NHS-ICs (p < 0.01). ATA-ICs [1.50 ± 0.24] versus medium [21.75 ± 6.24] (p = N.S.) and versus NHS-ICs [22.75 ± 7.59] (p = N.S.). ACA-ICs [1.80 ± 0.48] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). ARA-ICs [1.70 ± 0.14] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). Anti-Th/To-ICs [1.90 ± 0.42] versus medium (p < 0.01) versus NHS-ICs (p < 0.01). PAPS-ICs [2.02 ± 0.59] versus medium (p < 0.001) versus NHS-ICs (p < 0.01). SLE-ICs [1.42 ± 0.65] versus medium (p = N.S.) versus NHS-ICs (p = N.S.). NHS-ICs [1.08 ± 0.10] versus medium (p = N.S)