Figure 5. BALF- and lung epithelial cell-derived MVs contain pro-inflammatory miRNAs.

(A) Elevated miRNAs derived from BALF MVs after hyperoxia. The table was generated from miRNA microarray data as shown in Fig. 2A. (B,C) MVs were isolated from Beas2B cells (B) and primary epithelial cells (C). RNA was isolated from the MVs and quantified using the real-time quantitative RCR (qPCR). Individual miRNA expression levels were shown in bar graphs (D). Transwell migration assay of THP1 macrophages was performed after treated with mir-221 and/or mir-320a mimics which were transfected using lipofectamine 2000, as described in Material and Methods. (E–G) THP1 macrophages were treated with mir-221 and/or mir-320a mimics. After 16 h, MMP-9 levels (E) and TNF-α levels (F) from the culture medium were analyzed using gelatin zymography and ELISA, respectively. Western blot analysis was performed using total cell lysates with the indicated antibodies (G). For (B–F) data represent mean ± SD of three (D–F) or four (B,C) independent experiments. For (G) data represent two independent experiments with the similar results. Unprocessed original scans of gel and blots are shown in Suppl. Fig. 1.