Figure 2 of Nakamura, Mol Vis 2010; 16:425-437.


Figure 2. Analysis of protein expression from TRPM1 mutant minigenes. A: Schematic of TRPM1 minigenes. The minigene IVS2–3C>G contains an EGFP gene, exon 2, the exon 2 splice donor region, a portion of the β-globin (HBB) intron 2, the exon 3 splice acceptor region, and exon 3, and uses the CMV promoter and SV40 polyA site from the pEGFP-C3 vector. The minigene IVS8+3_6delAAGT contains an EGFP gene, exon 8, the exon 8 splice donor region, a portion of the HBB intron 2, the exon 9 splice acceptor region, and exon 9, and uses the CMV promoter and SV40 polyA site from the pEGFP-C2 vector. Mutations of IVS2–3C>G and IVS8+3_6delAAGT are shown. B-C: western blots of HEK293 cell extracts transfected with minigenes IVS2–3C>G (B) or IVS8+3_6delAAGT (C). WT represents protein from a wild-type minigene transfection, and asterisks represent spliced EGFP detected by an anti-Flag antibody. β-Gal and β-actin were used for transfection control and loading control, respectively.