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

Figure 5. Knockdown of BORIS resulted in decreasing expression of hTERT, stem cell and CSC markers genes.

Figure 5

NCCIT cells were engineered to stably exhibit knocked-down BORIS mRNA. BORIS sh-3, sh-4 and CTR sh (control with scrambled sequence) lentivirus were used to infect NCCIT cells. Each transduced cells were cultured with doxycycline to induce BORIS shRNA expression. Doxycycline-containing medium was replaced every 3 days. Each week over 1 month, mRNA levels of (A) BORIS, (B) CTCF and (C) hTERT were analyzed by qRT-PCR. Results were normalized with GAPDH and related to that of control cells (CTR sh) at each week. Error bars represent the mean ± SD of 3 independent experiments. (D) Telomerase activity was measured at each week by real-time quantitative PCR using TRAPEZE RT Telomerase Detection Kit. Values of telomerase activity of BORIS sh-3, sh-4 NCCIT-derived cells are related to that of control cells at each week. Error bars represent the mean ± SD of 3 independent experiments. (E) BORIS sh-3, sh-4 and CTR sh NCCIT-derived cells were cultured with doxycycline and after 7 days RNA was analyzed by qRT-PCR. mRNA levels of the indicated genes are related to that of control cells (CTR sh) after normalization with GAPDH. Error bars represent the mean ± SD of 3 independent experiments. Asterisks indicate statistically significant difference (p<0.05) between BORIS sh-3 or BORIS sh-4 and CTR sh cells.