Figure 1

MMGL isoform 4 interacts with the C1-C2 region of cMyBPC. A. Representative image of live cell fluorescence microscopy showing co-localization of cMyBPC and MMGL isoform 4. Each panel represents a single frame of the 25 images that were captured for the vertical Z-stack. The first four panels shows a single colour channel, while the image in the last panel shows an overlay of the four colour channels used. Column (iii) shows co-localization (yellow fluorescence) between dsRed-MMGL and GFP-cMyBPC, while column (iv) shows cardiac actin, a marker of the sarcomeric region. Scale bar: 0.02 mm. B. Representative image of live cell fluorescence microscopy showing that co-localization of MMGL isoform 4 and cMyBPC increases under adrenergic stress. Each panel represents a single frame of the 25 images that were captured for the vertical Z-stack. Each of the first three columns shows a single colour channel, while the image in the last column shows an overlay of the four colour channels used. Column (iii) shows co-localization (yellow fluorescence) between dsRed-MMGL and GFP-cMyBPC in the absence (-isopro) and presence (+isopro) of the beta-adrenergic agonist, isoproterenol. As evidenced by the increased yellow staining, co-localization levels between MMGL and cMyBPC increased ten minutes after the addition of isoproterenol. Scale bar: 0.02 mm. C. Quantification of co-localization shown in B demonstrates the significant increase in co-localization after the addition of isoproterenol (± SEM, *p < 0.05, n = 5). Change in co-localization was calculated using the CellR software and presented as a false colour image and percent co-localization as described by Loos et al., 2008 [29]. Abbreviations: isopro = isoproterenol