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. 2013 May 7;4:1822. doi: 10.1038/ncomms2791

Figure 3. praja2 inhibits the Hippo pathway and is required for GBM cell growth.

Figure 3

(a) HEK293 cells were transiently transfected with the indicated siRNAs and harvested at different confluence. Lysates were immunoblotted with the indicated antibodies. (b) Cumulative analysis of the experiments shown in a. (c) cells were transiently transfected with siRNAs and subjected to immunostaining with monoclonal anti-YAP. Images were collected and analysed by confocal microscopy. Scale bar, 20 μm. (d) Quantitative analysis (mean±s.e.m) of the experiments shown in (c), *P<0.01 versus control siRNA, Student’s t-test. (e) As in a, except the cells were transfected with flag-praja2rm or control CMV plasmid. (f) U87MG and FNP8 cells were transiently transfected with siRNAs and harvested at the indicated time points. Total RNA was extracted and subjected to RT–qPCR analysis for cyclin E (cycE) mRNA expression. Quantitative analysis is shown. These data are expressed as mean±s.e.m. of three independent experiments. *P<0.01 versus control siRNA (U87MG); #P<0.02 versus control siRNA (FNP8), Student’s t-test. (g) Growth curve of U87MG siRNA-transfected cells. Following transfection, cells were seeded in multiwell plates, harvested at the indicated time points and counted. A mean value ±s.e.m. of three experiments is shown. (h) Fluorescent-activated cell sorter analysis (FACS) of U87MG cells prepared as in c. The analysis was performed at 48 h following transfection. *P<0.05 versus control siRNA (G1); #P<0.05 versus control siRNA (G2/M).