Figure 4.

Expression of target gene candidates in response to induction of long non-coding RNA-Tcam1 (lncRNA-Tcam1) transcription by the Tet-off system. (A) Induction of lncRNA-Tcam1 transcription by the Tet-off system. GC-2spd(ts) cells were transfected with the construct for lncRNA-Tcam1 overexpression in response to the doxycycline (Dox) removal. After the selection of successfully transfected cells with puromycin, they were cultured in the presence (Dox+) or absence of Dox (Dox−) for 24 or 48 h. The cells were collected, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to measure the lncRNA-Tcam1 level using Gapdh as an internal control. The data represent the means ± SD from three independent experiments. A dramatic increase of lncRNA-Tcam1 transcription was observed in the Dox− sample at 48 h after the induction. (B) Expression of candidate genes at 48 h after the induction. qRT-PCR was performed, and the data were normalized to Gapdh. The values from Dox− samples were further normalized to those from Dox+ samples and expressed as fold-increases. The data represents the means ± SD from five independent experiments. The statistical significance was analyzed by Student’s t-test. *P < 0.05, **P < 0.01 compared to Dox+ samples.