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. 2005 Oct 6;102(42):15042–15047. doi: 10.1073/pnas.0507827102

Fig. 4.

Fig. 4.

The RS domain is not required to stimulate mRNA translation either in vivo or in vitro. (A) HeLa cells were transiently transfected with the pLCS-EDA reporter and either empty vector (control), wild-type SF2/ASF, or SF2/ASF cDNAs lacking RRM2 (RRM1/RS), RRM1 (RRM2/RS), or the RS domain (ΔRS), respectively (Left). Expression of the transiently expressed constructs was confirmed by Western blotting against the T7 epitope tag (Right). (B) Cytoplasmic extracts prepared from HeLa cells transfected with epitopetagged wild-type SF2/ASF or the ΔRS variant were fractionated across 10–50% sucrose gradients. Fractions were analyzed by Western blotting against the T7 epitope tag. (C) HeLa-cell-free in vitro translation assay comparing the ability of 5 pmol of exogenous SF2/ASF, ΔRS, or hnRNP A1 recombinant proteins to stimulate translation of in vitro-transcribed pLCS 3x EDA reporter mRNA.