Figure 2

Alternative splicing of Amer transcripts. RT-PCR of the N-terminus of Amer transcripts performed from E9.5 to E14.5 mouse embryos and HEK293T cells transfected with Wtx/Amer1 (HEK-A1), Amer2 (HEK-A2) or Amer3 (HEK-A3). Two RNA products are detectable by RT-PCR after transfection of HEK293T cells with mouse Wtx/Amer1 as previously reported [5] (right gel in the Wtx/Amer1 box). By contrast, a single band corresponding to the full-length transcript is detected in mouse embryos suggesting that regulation of splicing may be cell type dependent (left gel). For Amer2, only one transcript is detected endogenously in mouse embryos and after transfection of HEK293T cells. Nucleotide sequence analysis revealed a 357-bp (119 amino acids) deletion and indicated that Amer2 mRNA is produced by alternative splicing in the 5' end of exon 1. For Amer3, a single band corresponding to the full-length transcript is detected either in mouse embryos or after transfection of HEK293T cells. Results have been obtained with three different primer pairs for each Amer gene. RT: RNA samples incubated with reverse transcriptase; control: RNA samples incubated without reverse transcriptase as a DNA contamination control.