Figure 2.

IL10 Induces Physical Association of SHIP1 and STAT3

(A) J17 SHIP1^−/− cells expressing either His₆-SHIP1 or His₆-SHIP1 3PT were tested for their ability to be inhibited by IL10 in an LPS-stimulated TNFα production assay.

(B) J17 His₆-SHIP1 cells were stimulated with IL6, IL10, or ZPR-151 for 5 min His_6-SHIP1 was pulled down using Nickel beads and along with cell lysates probed with SHIP1, STAT3, and phospho-STAT3 antibodies.

(C) Single cell FRET analysis of J17 SHIP1^−/− cells expressing FRET pair fusion constructs, Clover-SHIP1 and mRuby2-STAT3, “mock” stimulated or stimulated with IL6, IL10, and ZPR-151 for 1 min. FRET efficiency was determined using the Acceptor Photobleaching method. Data represent % FRET efficiency of single cells from at least three independent experiments for each treatment (One-way ANOVA with Tukey's correction, ∗∗∗∗p < 0.0001).