Figure 3.

Time course of miR-29a release from HSC after TGFβ or PDGF-BB stimulation. Extracellular miR-29a levels were analyzed by real-time PCR (A and B), and LC3 protein immunofluorescence staining (C and D) was performed after 0, 4, 8, 12, 16, 20, and 24 hours of TGFβ (A and C) or PDGF-BB (B and D) treatment of HSC-T6 cells. Moreover, autophagosomes were stained by cationic amphiphilic tracer dye followed by fluorescence-activated cell sorter analysis (E). The percentages of autophagy-positive cells are presented in the bar diagram. (∗∗P < .001; data from 3 representative experiments are displayed) (F). Rapamycin and Torin-1 treatment for 24 hours were used as positive controls (G). Cy3-labeled miR-29a (red) was transfected, and colocalization with LC3 protein (green) expression was monitored after LC3 immunocytochemistry (scale bar = 15 μm) (H).