The Intronic Long Noncoding RNA ANRASSF1 Recruits PRC2 to the RASSF1A Promoter, Reducing the Expression of RASSF1A and Increasing Cell Proliferation (original) (raw)
Figure 4
Overexpression of ANRASSF1 increases cell growth and decreases UVC- and staurosporine-mediated cell death.
(A) Cell proliferation coefficients of _ANRASSF1_-transfected cells (pCEP4 ANRASSF1) relative to control cells (empty pCEP4), as measured using MTS assay. These data show the means ± SD from three independent experiments. *_t_-test p<0.03 relative to control. (B) Effect of ANRASSF1 overexpression on the cell population growth, as measured by counting the number of cells over time in culture and calculating the population doubling time. These data show the means ± SD from two independent experiments in triplicate. * _t_-test p<0.05. (C) DNA content histograms showing the effect of ANRASSF1 overexpression on the cell cycle at 48 h after exposure to UVC irradiation (40 J/m2). The cells were labeled with propidium iodide. Light gray indicates the sub-G1 population. (D) Percent of sub-G1 population from experiments identical to (C). These data show the means ± SD from three independent replicate experiments. *_t_-test p<0.05 relative to control. (E) Effect of ANRASSF1 overexpression on cell survival in the presence of the cytotoxic drug staurosporine (100 nM). The changes in impedance were measured using the xCELLigence system to continuously monitor cell attachment to the culture plates. Three independent biological replicates for either condition (pCEP4 ANRASSF1 or empty pCEP4) are shown, each representing the means ± SD of two or three technical replicates; the red lines show cells overexpressing ANRASSF1, and the blue lines show control cells carrying empty pCEP4) vector. *_t_-test p<0.001, for the cell indices at 170 h.