Fig. 2. FP-SMN is expressed at low levels relative to endogenous SMN, interacts with SIP1/Gemin2 and does not affect cell cycle. (A) Total cell lysates from the parental HeLa cell line, line CFP-SMNE8.8 and parental HeLa cells transiently transfected with a plasmid expressing CFP probed with anti-SMN and anti-FP antibodies. All three cell lines give a strong band at 38 kDa with anti-SMN, representing endogenous SMN. Line CFP-SMNE8.8 shows an additional, much weaker, band at 68 kDa, representing stably expressed CFP-SMN. A band of the same size is detected in line CFP-SMNE8.8 using the anti-FP antibody, whereas parental HeLa cells transiently transfected with CFP alone give a single band at 30 kDa. (B) Co-immunoprecipitations from total cell lysates of line GFP-SMNE10.3 using anti-FP antibodies. Detection of products using anti-SMN antibodies demonstrates the presence of GFP-SMN in the input material and the immunoprecipitated material (anti-GFP lane). A duplicate blot probed with anti-SIP1/Gemin2 shows a clear enrichment of SIP1/Gemin2 in the immunoprecipitated fraction (anti-GFP lane). (C) FACS analysis of synchronised cell populations stained with propidium iodide shows a similar profile for lines GFP-SMNE18.6, CFP-SMNE12A and the parental HeLa cell line.

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