Figure 4.

HA-Tagged PTMs Show Significant Maturation following Transfection in HEK293T

(A) In silico analysis of both genomes encoding either the 5′ PTM with NBD (PTM_NBD) or with CEP290 intron X-27 binding domain 07 (PTM_07) indicates an open reading frame (ORF) is present that is predicted to encode a 118.7-kDa peptide if the pre-mRNA matures without trans-splicing. (B) Representative western blot images of HA-tag and CEP290 using extracts from HEK293T harvested 72 hr following transfection with the indicated plasmids. HA-tagged CEP290 resulting from trans-splicing was expected at ∼290 kDa. α-tubulin was used as a loading control. (C) Densitometry quantification of CEP290 and the 118.7-kDa bands. Values per replicate group were standardized to α-tubulin and normalized to PTM_NBD. Error bars are SD; n = 6 independent experiments. p < 0.05 was considered significant.