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ICOS engagement alone activates Akt, ERK1/2, and p38 kinases. PBT lymphoblasts were rested overnight in the absence of IL-2 and then stimulated with microspheres coated with 5 μg/ml ICOSL.Ig for 1, 5, 15, 30, and 60 min. Cell lysates were analyzed for Akt, ERK1/2, and p38 phosphorylation (p-Akt, p-ERK1/2, p-p38, respectively) by immunoblot. Protein levels for total Akt, ERK1/2, and p38 expression were accessed as a loading control.
