Figure 5

Overexpression of eGFP-FMRP increased cell viability. Cells expressing either eGFP- or eGFP-FMRP were lysed with Trizol for RT-PCR (A) to confirm Fmr1 level. As a loading control, GAPDH level was also analyzed. (B) Similarly, protein level was checked by Western blot. (C) Either eGFP- or eGFP-FMRP expressing cells were treated with ETO (20 μM, 3 hr). Then, cells were analyzed by PI staining. Red fluorescence represents PI positive cells and green fluorescence represents GFP positive cells. White arrows indicate yellow spots which represent double positive for PI and GFP. (D) Graphs represent the quantification of the PI-positive cell number. (E) Treated cells were stained with TUNEL as described above. Similarly, red fluorescence represents TUNEL positive cells and green fluorescence represents GFP positive cells. White arrows indicate yellow spots which represent double positive for TUNEL and GFP. (F) Bar graph represents the quantification of the number of the apoptotic cells. * significantly different from non treated cells and ^# significantly different as compared with eGFP transfected cells (p < 0.01, n = 4). eGFP denotes HeLa cells transfected with eGFP-empty vector, eGFP-FMRP indicates cells transfected with eGFP-FMRP vector.