Additional file 1: Figure S1. of Regulation of IL-8 gene expression in gliomas by microRNA miR-93

Effects of pre-miR-93 and antagomiR-93 on morphology and apoptosis. A. Morphology of U251 glioma cells treated for 24 h with control, pre-miR-93 and antagomiR-93 molecules (200 nM). B,C. Effects of the different treatments on apoptosis on U251 and T98G glioma cell lines, as indicated. Apoptosis was analyzed by the Annexin-V release test [60] (B) or by caspase-3/7 production [61] (C). In panel B, the effects of a positive antagomiR-221 is also shown (see Brognara et al. (2014) [62]. Data represent the average S.D. of three independent experiments. ** = p < 0.01. (−): untreated cellular samples. Figure S2. Release of IL-8, VEGF, and MCP-1 by U251 glioma cells. U251 glioma cells were cultured for 48 and 72 h and protein release quantified by Bio-plex analysis. Data represent the average S.D. of three independent experiments. * = p < 0.05; ** = p < 0.01. Additional methods. Apoptosis was analyzed on U251 and T98G glioma cell lines after 48 h of treatment with pre-miR-93 or antagomiR-93 (200 nM). Cells were washed with sterile PBS (Phosphatebuffered Saline) and then tested with the Muse Annexin V Dead Cell kit (Millipore Corporation, Billerica, MA, USA) or Muse Caspase 3/7 kit (Millipore) [63]. The assays were performed with Muse Instrument (Millipore) [63], according to the instructions provided by the manufacturer. (PDF 360 kb)