Figure 5

Polycistronic expression using the 2 A linker sequence. (A) Grayscale image of palmTagRFP expression in a heart transfected with ITR-CAG-palmTagRFP-2A-h2bEGFP-ITR. (B) Grayscale image of h2bEGFP expression in heart from (A). (C) Merged channels from (A) and (B). (D) cropped region from (C). (E) Three-dimensional reconstruction of ventricular myocytes transfected with the construct from (A–D) imaged in situ. Scalebar = 20 um. (F) Three-dimensional reconstruction of an optically bisected ventricular myocyte transfected with ITR-CAG-palmTagRFP-2A-KDEL-EGFP-2A-h2bTagBFP-ITR imaged in situ. Scalebar = 10 um. (G) Three dimensional and volumetric reconstructions of E4 and E9 atrial myocytes transfected with the construct from (A–D) imaged in situ. Scalebar = 20 um. (H) Comparison of endoplasmic reticulum/sarcoplasmic reticulum distribution in ventricular myocytes transfected with the construct from (F) across stages (E4–E9) and in vivo vs in vitro. Scalebar = 20 um.