Fig. 5

LINC-PINT represses the expression of an invasion signature and induces CTNNB1 translocation. a Biological functions associated with genes differentially expressed upon LINC-PINT overexpression in HCT116 cells. b Heatmap representation of genes differentially expressed (DE) in HCT116 overexpressing LINC-PINT vs. HCT116 CTRL cells, involved in tumor cell adhesion, as defined by IPA (green, downregulation; red, upregulation). c Connection between CTNNB1 and genes regulated by LINC-PINT involved in cell movement and proliferation as predicted by IPA. d Immunoflorescence images of CTNNB1 (green) and DRAQ5 (blue, nuclear specific marker) in control cells (CTRL) and LINC-PINT overexpressing HCT116 cells (LINC-PINT). Scale bars: 20 μm (left). The fluorescence intensities of CTNNB1 are quantified by tracing a scanning line of 5 μm across the plasma membrane of the cell (right). e Subcellular fractionation and western blot analysis performed in HCT116. Three different fractions are loaded; total cell fraction (T), cytoplasmic fraction (C), and nuclear fraction (N) and probed for CTNNB1 and EGR1. GAPDH was used as cytoplasmic marker and LAMININ A/C as nuclear marker. f EGR1 overexpression restores invasive capacity of LINC-PINT overexpressing A549 and HCT116. Cells were either transduced with an empty vector (CTRL) or with LIC-PINT (LINC-PINT) and then transiently transfected to overexpress EGR1 (CTRL + EGR1 or LINC-PINT + EGR1). Data are from three biological replicates represented as mean ± SD of the fold change of invading cells. Significance was determined by one tail t-test (*P < 0.05, **P < 0.01, ***P < 0.001)