Fig. 7

miR-6794-5p induces macrophage M2 polarization via JAK1/STAT3 pathway. A Exosomes were isolated from the conditioned media of U251 cells overexpressing Bcl-w. After treatment of THP-1-derived macrophages with exosomes, the mRNA levels of CD163, CD206, and CD11b were measured by qRT-PCR analysis. B, C After THP-1-derived macrophages were treated with conditioned media from U251 and A549 cells transfected with miR-6794-5p mimics, respectively, mRNA levels of macrophage M2 markers (CD163, CD206, and CD11b) were measured by qRT-PCR analysis. D, E mRNA (D) or protein (E) levels of macrophage M2 markers (CD163, CD206, and CD11b) in THP-1-derived macrophages overexpressing the miR-6794-5p mimic were verified by qRT-PCR or Western blot analysis, respectively. F Using the TCGA dataset, expression of CD206 was compared with normal (n = 5) and GBM patients (n = 167) and displayed as box and whisker plots. G The expression level of CD206 in the lung tissues at each stage of lymph node metastasis of patients with lung cancer was confirmed by IHC. H Kaplan-Meier overall survival curves of patients with lung squamous cell carcinoma according to the expression of CD163, CD206 (MRC1), and CD11b (ITGAM). I, J Expressions of p-STAT3, STAT3, p-JAK1, JAK1, and SOCS1 in miR-6794-5p-overexpressed (I) or SOCS1-knockdown (J) THP-1-derived macrophages were shown by Western blot analysis. K Expression of STAT3 protein after transfection with STAT3 against siRNA in macrophages overexpressing miR-6794-5p (left). Expression mRNA levels of macrophage M2 markers (CD163, CD206, and CD11b) and IL-10 were checked in the indicated cells (right). L After overexpressing empty vector, miR-6794-5p and miR-6794-5p + SOCS1 in THP-1-derived macrophages, the expression of SOCS1 protein was confirmed by Western blot analysis (left), and the mRNA expression levels of macrophage M2 markers (CD163, CD206, and CD11b) were confirmed by qRT-PCR (right) in indicated cells. β-actin was used for normalization in Western blot analysis. The values were normalized to GAPDH in qRT-PCR. The experiment was repeated with triplicates and representative Western blotting images are shown. The data are presented as the mean ± S.D. after triplicate. *P < 0.05; **P < 0.01; ***P < 0.001. A-D Student’s t-test. K, L One-way ANOVA followed by bonferroni comparison test