Fig. 3

Silencing of selected lncRNAs in the SNB19 GBM cell line leads to reduced growth and colony formation. (a) Relative expression levels of TCONS_l2_00001282, lnc-GBMT-6, and lnc-NBN-1 after targeted or control siRNA transfection in the SNB19 cell line. All of the lncRNAs presented reduced expression after siRNA transfection. The results show the mean expression difference together with the standard deviation from measurements representing three separate biological experiments (separate wells on a well-plate), each with three technical replicates (separate measurements from one well). (b) Alamar Blue assay showing reduced cell growth after the silencing of lncRNAs TCONS_l2_00001282, lnc-GBMT-6, and lnc-NBN-1. The figure shows the mean relative growth from six biological replicates (separate wells on a well-plate) with the standard deviation after five days in comparison with that on day zero. ****p < 0.0001, one-way ANOVA. From each biological replicate, six technical replicates (separate measurements from one well) were taken for the Alamar Blue assay. The results for similar, significantly reduced cell growth were acquired in two separate experiments in the case of TCONS_l2_00001282 and in three separate experiments for lnc-GBMT-6 and lnc-NBN-1. (c) Representative images of cell culture dishes after the clonogenic assay for TCONS_l2_00001282, lnc-GBMT-6, and lnc-NBN-1 lncRNAs that had growth effects in the Alamar Blue assay. d) Cell colony formation was reduced after the silencing of TCONS_l2_00001282, lnc-GBMT-6, or lnc-NBN-1 lncRNAs in the SNB19 cell line. The figure shows the mean number of colonies from three biological replicates (separate wells on a well-plate) together with the standard deviation after transfection with control siRNA or siRNAs against each lncRNA. **p < 0.01, ***p < 0.001, one-way ANOVA. Similarly, significantly reduced colony formation was detected in two separate experiments after these lncRNAs were silenced.