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. 2013 Apr 25;8(4):e62305. doi: 10.1371/journal.pone.0062305

Figure 1. Procedure for establishment of human protein replacement stable cell line using snoMEN.

Figure 1

(A) Structures for targeted endogenous SMN1/UBF1 protein replacement plasmids (pGFP–SMN1snoMENv1-PR/pmCherry–UBF1snoMENv1-PR). These constructs have three snoMEN sequences as previously described [1], except that the M box sequences are complementary to endogenous SMN1/UBF1 pre-mRNA sequences (See Materials and Methods). (B) Procedure of stable cell line establishment. Transfected cells were selected under G418 treatment as previously described (http://www.lamondlab.com/f7protocols.htm). Cells were cultured for at least 14 passages before analysis to confirm stable FP–protein and snoMEN expression. (C) Images of protein replacement stable cell lines. Expression of FP proteins was confirmed by fluorescence imaging. Bar length is 10 µm.