Figure 2. The lncRNA LUST overexpression is an early event in the spheres formation process.
A) LUST, CD24 and CD44 mRNA levels measured by qRT-PCR in HT-29 derived spheres formed with colon carcinoma cells, respectively after 14 and 35 days cultured in ultra-low attachment conditions and in FBS-induced differentiation cells. HPRT transcript was used as housekeeping control gene for normalization. Error bars indicate the standard error of the mean (SEM). *p≤0.05; B) FACS analysis of spheres formed with colon adenocarcinoma cell lines (upper panels) and FBS-induced differentiated cells (lower panels) showing corresponding loss of expression of stem-like markers (blue population). Histograms report the mean fluorescence intensity (MFI, range 0–10,000) of CD24 (FITC) and CD44 (PE-Cy7); C) Representative images of HT-29-derived spheres formed with colon adenocarcinoma cells with characteristic spheroid morphology (top) and differentiated spheres showing adherent morphology upon FBS-induced differentiation (bottom).