Phospho p38mapk t180 y182

All antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA) and used as follows: BAX (no. ab32503, diluted: 1:1000), BCL-2 (no. ab32124, diluted: 1:1000), CASPASE8 (no. ab32397, diluted: 1:1000), PARP1 (no. ab32138, diluted: 1:1000), GADD45A (no. ab203090, diluted: 1:500), p38/MAPK (no. ab170099, diluted:1:1000), phospho-p38MAPK (T180 + Y182) (no. ab195049, diluted: 1:1000), ERK1/2 (no. ab184699, diluted: 1:1000), phospho-ERK 1/2 (T202 + Y204) (no. ab223500, diluted: 1:1000) were purchased from Abcam (Cambridge, USA). Cleaved-caspase 8 (no. 8592, diluted: 1:1000), GAPDH (no. 5174), CDC2 (no. 28439, diluted: 1:1000), CYCLINB1 (no. 4138, diluted: 1:1000), PLK1 (no. 4535, diluted: 1:1000), MMP-1 (no. ab137332, diluted: 1:1000), MMP-9 (no. 13667s, diluted: 1:1000), VEGF-1 (no. 2893s, diluted: 1:1000). Anti-rabbit and anti-mouse LgG (H + L) (Dylight (TM) 800 4×PEG Conjugate) secondary antibodies used in this study were purchased from Cell Signaling Technology and used at a 1:30000 dilution in the experiments.

Phospho-p38 MAPK (T180/Y182) + Total In-Cell ELISA Kit (Abcam, Cambridge, MA) was used to measure phosphorylation activity of p38 MAPK per manufacturer’s instructions. The effect of MAP infection on the ratio of p-p38 MAPK/p38 MAPK in the presence of different levels of 5-HT (0, 100, 250, 500 ng/mL) was presented with or without 1 uM of p38 MAPK inhibitor (SB220025) following 24 h of incubation.