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[Preprint]. 2024 Oct 25:2024.10.22.619581. [Version 1] doi: 10.1101/2024.10.22.619581

Fig. 3. Canonical and alternative transcripts specify CDS or iORF expression, respectively.

Fig. 3

a, GENCODE canonical (C) and alternative (Alt) transcripts from four representative genes were cloned from HEK 293 cell cDNA from the 5’ end through the iORF stop codon, with an HA-FLAG epitope tag appended to the 3’ end of the iORF. Anti-FLAG Western blotting was performed after transient transfection to probe iORF expression from each context. b, The start codon of CDSs was mutated to TAA within canonical transcripts to test its suppression of iORF expression. Western blotting for an epitope tag appended to the 3’ end of the iORF was performed after transient transfection.c, GENCODE C and Alt transcripts were cloned from HEK 293 cDNA from the 5’ end through the CDS stop codon, immediately before which an HA epitope tag was encoded. Western blotting from transiently transfected cells was performed to examine CDS expression from each context.