AS1517499, a STAT6 inhibitor, impacted Meta1 gastroid growth and gene expression. (A) Time-course diagram of the treatment to Meta1 gastroids with rIL-13 and/or AS1517499. (B) Representative bright field images of Matrigel domes containing Meta1 gastroids at different times of treatment. Quantitation of gastroid diameters showed a significant difference in Meta1 gastroids treated with AS1517499. Scale bar: 500 μm. Mean ± SD (N = 3 independent experiments). ∗P < .05, ∗∗∗P < .001, ∗∗∗∗P < .0001. (C) Quantitative RT-PCR showing relative expression levels of SPEM-related genes (Muc6, Tff2, Gkn3, Cd44v9, Atf3, and He4). Mean ± SD (N = 3 independent experiments). ∗P < .05, ∗∗P < .01, ∗∗∗P < .001. (D) Immunoblotting shows significant down-regulation of phosphorylated STAT6 and ATF3 in Meta1 gastroids treated with STAT6 inhibitor (AS). Mean ± SD (N = 4 independent experiments). ∗∗∗P < .001.