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. 2017 Oct 23;13(10):e1006690. doi: 10.1371/journal.ppat.1006690

Fig 4. Down-regulation of ERK1/2 and p38 activation in ILC3s by lethal toxin.

Fig 4

MNK-3 cells were pretreated with lethal toxin (1 μg/ml) for 2 hr followed by IL-23 stimulation as indicated. Cell lysates were harvested and subjected to immunoblot analysis for MEK1, MEK2, MKK3, MKK6, phosphorylated ERK1/2 (p-ERK1/2), total ERK1/2 (t-ERK1/2), phosphorylated p38 (p-p38), total p38 (t-p38) and actin. Shown is a representative blot for n = 2–4 experiments. (B) MNK-3 cells were pretreated with anti-IL-23 p19 antibody (5 or 50 μg/ml) for 1 hr followed by IL-23 stimulation for 15 or 30 min. Cell lysates were subjected to immunoblot analysis for phosphorylated ERK1/2 (p-ERK1/2), total ERK1/2, phosphorylated p38 (p-p38), total p38 and actin. Shown is a representative blot for 2–4 experiments. (C) In vitro expanded human Lin- CD127+ cells or (D) mouse Lin- CD127+ c-Kit+ Thy1.2+ cells expanded from splenocytes of Rag1-/- mice were treated with lethal toxin for 2 hr followed by IL-23 stimulation for 30 mins. Cell lysates were analyzed for MEK1, phosphorylated ERK1/2 (p-ERK1/2), total ERK1/2 and actin by immunoblot analysis. Shown are representative blots from 2 independent experiments for both C and D.