fig2

Figure 2. Content of microRNA miR-221-3p and miR-222-3p in HT-29 and LoVo cells, HT-29 and LoVo tumors and plasma isolated from HT-29 and LoVo xenotransplanted mice. A: representative Reverse Transcription droplet digital PCR plots performed using RNA isolated from tumor cells, plasma from tumor-free mice, and plasma from xenografted mice; B, C: content of miR-221-3p and miR-222-3p in cells, xenotransplanted tumors and plasma. RNA was extracted from frozen cell (5 × 10⁵) pellets by the TRI-Reagent (Sigma-Aldrich, St.Louis, MO, USA), the procedure is described in Gasparello et al.^[55]. All RNAs were stored at -80 °C until the use. Reverse Transcription droplet digital PCR assays for microRNA expression analysis were performed to quantify the levels of miR-221-3p and miR-222-3p. 300 ng of total RNA (from cells and tissues) and the RNA isolated from 150 µL of plasma were reverse transcribed and analyzed for miR-221-3p and miR-222-3p as described by Gasparello et al.^[55]