P44 42 mapk erk1 2 sirna

After a 2-day culture, PANC1 cells were reseeded into six-well plates at 2 × 10⁵ cells/well. After 24 h, the Rab43 siRNA (50 pmol, Santa Cruz Biotechnology Inc., Dallas, TX, USA), p44/42 MAPK (ERK1/2) siRNA (100 nM) and Akt siRNA (100 nM, Cell Signaling Technology) were delivered to cells via a lipid-mediated Lipofectamine 2000 Transfection Reagent. A nonrelated scramble siRNA (Invitrogen) and SignalSilence control siRNA (Cell Signaling Technology) were used as a transfection control.

Most of the small interfering RNAs and controls were purchased from Santa Cruz Biotechnology. They are TNF-α siRNA (sc-37216), ATM siRNA (sc-29761), NF-κB p65 siRNA (sc-29410), hMMP3 siRNA (sc-29399), hMMP9 siRNA (sc-29400), hMMP13 siRNA (sc-41559) and control siRNA (sc-37007). p38 MAPK siRNA (#6564) and p44/42 MAPK (Erk1/2) siRNA (#6560) were obtained from Cell Signaling Technology. For siRNA transfection, 30-50% confluent cells were transfected with siRNA using Lipofectamine 2000. The cells were harvested 48 h after transfection. The final concentration for siRNA is 100 nM. The silence effects of indicative siRNA in NCI-H446 cells were validated in Supplementary Figure S8.