Fig. 4

Unsupervised hierarchical cluster analysis of miRNA expression in LPS pre-Exo uncovers let-7b as a regulator of TLR4 in macrophage polarization. a miRNA expression profiling on total RNA isolated from un-Exo (n = 3) and LPS pre-Exo (n = 3). A heatmap was generated after supervised hierarchical cluster analysis. Differential miRNA expression is shown by red (upregulation) versus green (downregulation) intensity (LPS pre-Exo versus un-Exo, twofold change, p < 0.05). b Validation of unique miRNAs in LPS pre-Exo using real-time RT-PCR. c Real-time RT-PCR analysis of let-7b expression in treated THP-1 cells at 48 h. Data are presented as the mean ± SEM of three separate experiments. *Compared with Con, p < 0.05; ^#compared with HG, p < 0.05. d Immunofluorescence staining displayed TLR4 activity in treated THP-1 cells at 48 h