The SARAH Domain of RASSF1A and Its Tumor Suppressor Function
Figure 2
Proliferation analysis of RASSF1AΔSARAH expressing lung cancer cells. (a) A549 lung cancer cells were transfected with pCMV-Tag1, RASSF1A, RASSF1C, and RASSF1AΔSARAH (RASSF1AΔS) and stable clones were analyzed. RASSF1 expression of clones was confirmed by RT-PCR using a Flag-specific forward primer and primer L27111. Products (RASSF1A and RASSF1AΔS, 585 bp; RASSF1C, 374 bp) were analysed with controls (pCMV-Tag1 and H2O) and a 100 bp ladder (M) on a 2% Tris-borate EDTA agarose gel. (b) Growth curve of A549 cells stably transfected with the indicated constructs. Clones were analyzed by seeding cells in 6-well plates. Every 24 hours, cells were counted using a Neubauer chamber. Three independent experiments were performed and the mean and standard deviation is plotted. (c) Colony sizes in a soft agar experiment after 4 weeks. Examples of colonies expressing pCMV-Tag1, RASSF1A, and RASSF1AΔS are shown. (d) 25 colonies were measured and the average colony size was calculated. Statistical significance -values are indicated.
