Skip to main content
. 2014 Aug 26;9(8):e104401. doi: 10.1371/journal.pone.0104401

Figure 5. Effect of [6]-gingerol on PMA-induced phosphorylation of MAP kinases and activation of AP-1 and NF-kappaB in SW-480 cells.

Figure 5

(A) Overnight grown SW-480 cells were treated with 50 ng/ml of PMA for different time intervals (0–120 min) and the whole cell lysate was immunoblotted onto PVDF membrane, probed using antibodies against phospho-ERK1/2, phospho-JNK and phospho-p38, developed by ECL. The kinetics of PMA-induced phosphorylation of these MAP kinases were followed from this blot (B) SW-480 cells were pre-treated with 200 µM [6]-gingerol before treating with 50 ng/ml of PMA for 30 min and the whole cell lysates were immunoblotted, probed and detected as above. The relative fold difference of bands with control treatments are indicated below each lane. β-actin served as the loading control in each case. (C) SW-480 cells, pre-treated with indicated concentrations of [6]-gingerol for 2 h, were treated with PMA (50 ng/ml) for 30 min. The nuclear extracts from each treatment were analysed for the activation of AP-1 or (D) NF-kappaB, by performing the transcriptional binding assay on isotope labelled AP-1/NF-kappaB specific DNA binding probe and analysing it on electrophoretic mobility shift assay (EMSA). The arrowhead in each case indicates complexes between AP-1/NF-kappaB and DNA probe.