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. 2014 Dec 10;9(12):e114573. doi: 10.1371/journal.pone.0114573

Figure 4. Berberine inhibits mTORC1 signaling and ERK activation in PDAC cells.

Figure 4

Cultures of MiaPaCa-2 (Panels A and B) or PANC-1 cells (panels C and D) were incubated in the absence or in the presence of increasing concentrations of berberine. Then, the cells were stimulated for 1 h with 5 nM neurotensin and 10 ng/ml insulin and lysed with 2X SDS-PAGE sample buffer. The samples were analyzed by SDS-PAGE and immunoblotting with antibodies that detect the phosphorylated state of S6K at Thr389, S6 at Ser240/244, and ERK at Thr202 and Tyr204. Immunoblotting with total S6K, S6 and ERK was used to verify equal gel loading. The quantification of the immune signals was performed using Multi Gauge V3.0. The results are presented in the plots shown in panels B and D. The values represent the mean ± SEM (n = 3) of S6K, S6 and ERK phosphorylation expressed as a percentage of the maximal response obtained in 3 independent experiments.