Figure 3
Functional effects of LINC00958 or LINC01296 knock-down in FL3 cells. FL3 cells were transfected either with a non-targeting LNATMGapmeR (Scr) or 2 different LNATMGapmeR separately targeting LINC00958 or LINC01296 (LNA-1 and LNA-2) at the indicated concentrations (10, 25 or 50 nM). (a) Dose-dependent knock-down efficiency was verified by RT-qPCR 48 hours post-transfection. Results were normalized to Scr. (b) Dose-dependent viability was assessed by MTT reduction 24, 48 and 72 h post-transfection and expressed as percentage compared to viability of untransfected cells. (c) Dose-dependent cell death was determined by the LDH release assay 24, 48 and 72 h post-transfection and depicted as percentage of released LDH out of total cellular LDH. (d) Cell morphology was examined upon candidate knock-down by phase contrast microscopy 72 h post-transfection (25x magnification, scale bar 40 μm). Cellular migration (e) and invasion (f) of transfected FL3 cells (25 nM LNATMGapmeR). (g) Anoikis resistance upon candidate knock-down (25 nM LNATMGapmeR). Cell viability was assessed by both automatic and manual cell counting. Results were normalized to Scr. FCS: Fetal calf Serum. Columns in (b,c and g) represents the average of two or three independent experiments, each measured in three to five replicates. Bars: SD and *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.
