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SC_12-0166_sm_supplFigure1.tif4135KSupplemental Figure 1. (A) qRT-PCR analysis (left panel) of fibroblasts transfected with pCMV-Sport 6 plasmid carrying the human CD29 (fibr-CD29) full length cDNA or of fibroblasts electroporated in the absence of DNA as control (fibr-CTR). Data were normalized to 1 for fibr-CTR and to GAPDH mRNA. * P < 0.01 vs fibr-CTR. Right panel, Western Blot analysis of CD29 in fibr-CTR, in MVs derived from fibr-CTR, in fibr-CD29 and in MVs derived from fibr-CD29. CD29 transfected fibroblasts released MVs expressing CD29. (B) Nanoparticle tracking analysis (NTA) showing that the RNase treatment (right panel) did not affect MV morphology and size. Left panel shows untreated MV-HLSC. Curve 1 describes the relationship between particle number distribution (left Y axis) and particle size (X axis); curve 2 shows cumulative percentage distribution of particles (percentile in right Y axis) and particle size (X axis). Mean size and particle concentration values were calculated by the NTA software that allow analysis of video images of the particle movement under Brownian motion captured by Nanosight LM10 and calculation of the diffusion coefficient, sphere equivalent, and hydrodynamic radius of particles by using the Strokes-Einstein equation. (C) Measure of selected miRNAs in MVHLSC treated 5U/ml (white bar) of RNase in respect to control MV-HLSC (black bar) expressed as 2-(ΔCt) as described in Methods. Data were normalized to 1 for MV-HLSC and expressed as mean ± SD of 4 different experiments. * P < 0.01 vs MV-HLSC. (D) Measure in MV-HLSC of the levels of different miRNAs relative to the small nucleolar RNA, RNU48 expressed as Log 2-(ΔCt).
SC_12-0166_sm_supplFigure2.pdf911KSupplemental Figure 2. Evaluation of in vivo tumor proliferation and apoptosis (A) Representative micrographs of PCNA staining of HepG2 tumors. Scale bar, 50 μm. (B) Representative micrographs of Tunel staining. Scale bar, 50 μm.
SC_12-0166_sm_supplFigure3.tif344KSupplemental Figure 3. Comparative expression of specific miRNAs and correspondent targets between HepG2 and normal human hepatocytes. (A) Log of relative expression of miR451, miR223, miR31 and miR21 in normal human hepatocytes (Hep; white bar) in respect to HepG2, evaluated by qRT-PCR. (B) qRTPCR analysis of miR451 and miR31 correspondent targets in HepG2 (grey bar) and in Hep (white bar). Data (A and B) were normalized to 18S ribosomal 1 mRNA and to 1 for HepG2 and expressed as mean ± SD of 4 different experiments. * P < 0.05 vs HepG2.
SC_12-0166_sm_supplTable1.pdf60KSupplementary Table 1

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