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. 2014 Oct 3;9(10):e109921. doi: 10.1371/journal.pone.0109921

Figure 6. BORIS knockdown impairs cell senescence but not apoptosis.

Figure 6

(A) Cell proliferation over 1 moth of dox-induced BORIS shRNA cells were analyzed by MTT assay. Results of the two specific BORIS-shRNA (BORIS sh-3 and sh-4) NCCIT-derived cells are indicated as a percentage compared to the cell proliferation of control cells (scrambled shRNA, CTR sh). Error bars represent the mean ± SD of 3 independent experiments. (B) After dox-induction of the BORIS specific shRNA in NCCIT cells, apoptosis was tested at each week using Annexin V Apoptosis Detection Kit. Results show the percentage of apoptotic cells (late apoptotic AnnexinV+/7AAD+ and early apoptotic AnnexinV+/7AAD−) of BORIS sh-3 and sh-4 cells compared to the control cells. Error bars represent the mean ± SD of 3 experiments. (C) The senescence-associated β-galactosidase (SA-β-gal) staining was performed using β-galactosidase staining kit. SA-β-gal was analyzed after 2 and 4 weeks of dox-induction of the BORIS specific shRNA in NCCIT cells. Results show the percentage of senescent cells of BORIS sh-3, BORIS sh4 and control cells. Error bars represent the mean ± SD of 3 experiments. Asterisks indicate statistically significant difference (p<0.05) between BORIS sh-3 or BORIS sh-4 and CTR sh cells. Representative images of NCCIT cells after 4 weeks of BORIS knockdown and stained with SA-β-gal were shown.