Figure 5
NET-mediated transfer of miRNA-142-3p to macrophages. (A) PMA-differentiated THP-1 macrophages were exposed for 2 h to PMA-induced NET, following washing and complete RNA extraction for hsa-miR-142-3p (left group) and hsa-pri-miR-142 (right group) expression evaluated by quantitative PCR using Taqman probes. (B–E) PMA-differentiated THP-1 macrophages were exposed for 2 h to PMA-induced NET associated with specific hsa-miRNA-142-3p inhibitor or scramble sequence control, or to the hsa-miRNA-142-3p inhibitor or the scramble sequence control alone. After, cells were washed, and complete culture medium was added. Two hours (B,C) or 18 h later (D,E), cells were analyzed through flow cytometry for detection of hsa-miRNA-142-3p inhibitor or scramble sequence control in cells; both being labeled with the TYE™-665 fluorochrome. Data are shown as ratio of mean fluorescence intensity (MFI). Results in (A) are from 5 independent experiments using paired Student’s t test, and in (B,D) are from 4 independent experiments using one-way ANOVA, with Bonferroni post-hoc test, displaying mean ± standard error of mean of the test group. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.
