Fig. 3

miR-326 over-expression exerts tumour suppressive effects in glioblastoma. a. Log₂ transformed miR-326 expression value obtained from qRT-PCR analysis upon its over-expression in LN229 cell line. b, c. LN229 cells were transfected with Pre-miR-neg or Pre-miR-326, cells were counted and indicated numbers of cells (a = 3000 and b = 6000 cells) were plated in duplicates in a 12-well cluster plate. After 3 weeks of plating, the colonies were stained with crystal violet, photographed (b) and counted (c). % colony density and ± Sd is plotted. d. LN229 cells transfected with Pre-miR-neg or Pre-miR-326 were subjected to proliferation assay. Proliferation was assessed by MTT assay, measuring OD (570 nm) for both miR-neg and miR-326 at each time point. For each condition and time point, the samples were plated in triplicates and the standard deviation was calculated over three independent biological replicates. Significance is indicated for different time points. e, f. LN229 cells transfected with Pre-miR-neg or Pre-miR-326 were subjected to scratch assay (e), migratory potential of cells was quantitated by the percent of wound remaining after the indicated time point (f). p values were calculated by student’s t-test using graph pad prism and the symbols are indicated. (*) p ≤ 0.05; (**) p ≤ 0.01; (***) p ≤ 0.001