miR-374a-5p regulated key monocyte functions such as migration and T-cell activation. (A) Migration assays of monocytes from seven HC and six IBD patients transfected with SCR or miR-374a-5p mimic. Cells were monitored with Calcein, and a ChemoTx Disposable Chemotaxis System was used, with CCL2 as a chemoattractant. Mean and SD are shown. (B) Migration assays of CD4⁺ T cells, using as chemoattractant the supernatant of monocytes transfected with SCR or miR-374a-5p mimic from LPS-stimulated HC (left) or IBD patients (right). (C) Experimental design chart of coculture experiments between anti-CD3–stimulated CD4⁺ T cells and autologous LPS-stimulated monocytes pretransfected with SCR or miR-374a-5p mimic. (D) T cell activation markers (CD25 and HLA-DR) evaluated by flow cytometry on CD4⁺ T cells after coculture. (E) Representative dot plot showing the differentiation status of CD4⁺ T cells from HC before coculture (left). The percentages of T_CM, T_N, T_EM, and T_EMRA were analyzed by flow cytometry after 24 h of coculture (right). Mean and SD of five independent experiments are shown. SCR, scrambled control; T_CM, CD45RA⁻CD62L⁺; T_N, CD45RA⁺CD62L⁺; T_EM, CD45RA⁻CD62L⁻; T_EMRA, CD45RA⁺CD62L⁻. *, P < 0.05. Wilcoxon test. Mann–Whitney U test was used in A between HC and IBD.