The clinical relevance of macrophage-expressed Shp2 in IBD patients. (A) The expression of Shp2 in CD68⁺ colonic macrophages were examined in mucosal biopsy specimens by immunofluorescence staining. Bars, 20 µm. Data are representative of three independent experiments. (B) The percentage of CD68⁺Shp2^hi cells was quantified. Each group contained three individuals. Data are mean ± SEM and are compiled from three independent experiments. (C) The expression level of Shp2 in peripheral monocytes was evaluated by qPCR. (D) Association between mRNA levels of Shp2 and TNF-α or IL-6 was analyzed by Spearman’s rank correlation test. Data are mean ± SEM and are from one independent experiment. (E–H) THP-1 monocytes (E and F) or THP-1–differentiated macrophages (G and H) were transduced with Shp2-knockdown lentivirus (Lv-Shp2) or control lentivirus (Lv-scr), followed by stimulation with IL-10 for 30 min. The phosphorylation of STAT3 was determined by Western blot (E and G). IL-10 inhibition of TNF-α was measured by ELISA (F and H). (I) GST-tagged STAT3 and myc-tagged Shp2 full length or truncations were overexpressed in HEK293T cells. The interaction between STAT3 and various Shp2 domains was detected by co-IP. (J and K) THP-1–differentiated macrophages were stimulated with TNF-α (20 ng/ml); the level of Shp2 was evaluated by Western blot (J) or flow cytometry (K). Data are mean ± SEM and are representative of at least two independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; two-tailed unpaired Student’s t test (B, C, F, H, and K) and Spearman’s rank correlation test (D).