Fig. 4: In vivo expression and tissue distribution of intramyocardial injected modRNA of the intrabodies VHH2 B4B4-HA and VHH2 C6C6-HA.
From: Blocking phospholamban with VHH intrabodies enhances contractility and relaxation in heart failure
a Overview of the experimental procedure: Mice (n = 12) were injected intramyocardially with modRNA, sacrificed after 24 or 72 h; hearts were collected for western blot analysis (n = 3 at 24 h, n = 3 at 72 h); and immunofluorescence imaging (IF) of tissue sections (n = 3 at 24 h, n = 3 at 72 h). 6 mice, not treated with modRNA were used as control samples: western blot (n = 2 at 24 h; n = 2 at 72 h) and IF (n = 1 at 24 h; n = 1 at 72 h) b, c Western blot analysis of whole heart lysates (VHH2 B4B4: n = 3 at 24 h, n = 3 at 72 h; VHH2 C6C6-HA: n = 3 at 24 h, n = 3 at 72 h; control: n = 1 at 24 h, n = 1 at 72 h). The membranes were stained with anti-HA to reveal the expression of the HA tagged intrabodies: b VHH2 B4B4-HA, c VHH2 C6C6-HA. An anti-cofilin antibody was used as a loading control. Lane 1 contains a molecular weight marker (M) and sizes of the bands (in kDa) are indicated on the left. Western blots were performed n = 3 times with similar results. d Immunofluorescence imaging of transfected mouse hearts after 24 h of in vivo expression of VHH2 B4B4-HA (top panels) and VHH2 C6C6-HA (bottom panels). Slices were stained with DAPI, the SERCA2a antibody (red) and the anti-HA antibody (green). Identical immunofluorescence staining was repeated twice for tissue sections of mice transfected with VHH2 B4B4-HA at the 24 h time point and repeated once for tissue sections of mice transfected with VHH2 C6C6-HA at the 24 h time point (using 10 slices per heart) and resulted in similar staining patterns. Source data are provided as a Source Data file.
