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. 2022 Dec 20;119(6):1361–1376. doi: 10.1093/cvr/cvac191

Figure 2.

Figure 2

The CARMEN-201 Exon 2 contains a functional transposable element implicated in SMC specification. (A) Expression of CARMEN isoforms and (B) SMC markers (MYH11; CNN1; TAGLN; CALD1) in differentiating fetal cells either untransfected (None), transfected with the SAM system in the absence of gRNA (SAM) or with the SAM system with a gRNA targeting sequences upstream the CARMEN TSS (SAM/gRNA). (C) Representative images and quantification of SMCs in cultures of differentiating fetal CPCs transfected as in (A). Scale bar: 50 µm. (D) Position of the MIRc transposable element in the CARMEN-201second exon, and sequence conservation. (E) Expression of CARMEN-201 using either a primer pair specific for the endogenous transcript (P1) or the exogenous Exon 2 (P3), and (F) SMC markers (MYH11; CNN1; TAGLN; CALD1) in differentiating fetal CPCs either not transduced (None), transduced with a lentiviral vector encoding CARMEN(C)-201 Ex2 or transduced with a lentiviral vector encoding a mutated C-201 Ex2 (C-201 mutEx2). (G) Representative images and quantification of SMMHC-positive CNN1-positive SMCs in cultures of differentiating fetal CPCs transfected as in (F). Scale bar: 50 µm. Data represent mean values ± SEM; *P < 0.05 as compared with fetal CPCs in expansion; §P < 0.05 compared with the indicated conditions (n = 3–6). ANOVA with post hoc Tukey. See also Supplementary material online, Figure S3 and S4.