DCA stimulation of EGFR-MAPK signaling in <i>ex vivo</i> human intestinal enteroids is mediated by PA.

(A) Human intestinal enteroids were pre-serum-starved before incubation with 5 μM DCA for various time points to ensure total serum starvation time is 2 hours for all conditions. Whole cell lysates were collected and blotted using antibodies against pMEK, total MEK, pERK, total ERK, pAkt or total Akt, as well as the loading control actin. (B) Human intestinal enteroids were serum-starved for 1 hour and 55 minutes before incubation with 1–200μM DCA for an additional 5 minutes. Whole cell lysates were collected and blotted using antibodies against pMEK, total MEK, pERK, total ERK, pAkt or total Akt, as well as the loading control actin. (C) Pre-serum-starved human intestinal enteroids were treated with 0.75μM FIPI for 25 minutes and subsequent co-incubation with both FIPI and 5μM DCA. Whole cell lysates were collected and blotted using antibodies against pMEK, total MEK, pERK, total ERK, pAkt or total Akt, as well as the loading control actin. In all signaling experiments, 3 individual experiments were performed separately and quantitation is shown as mean±SEM. Statistical significance was evaluated using one-way ANOVA, with * indicating p<0.05.