The turnover rate of α-actinin assessed in a pulse-chase experiment. Xiaohu Fan Bryan G. Hughes Mohammad A. M. Ali Woo Jung Cho Waleska Lopez Richard Schulz 10.1371/journal.pone.0129176.g002 https://plos.figshare.com/articles/figure/_The_turnover_rate_of_945_actinin_assessed_in_a_pulse_chase_experiment_/1449421 <p>NRVM were transduced with lentiviral vector harboring C-terminal HaloTag labeled α-actinin. 48 hr after transduction, cells were stained with excess TMRDirect (red fluorescence), a cell permeable ligand which forms a stable covalent bond with HaloTag α-actinin. At specified time points after TMRDirect staining as indicated, a different Halo-tag ligand (R110Direct, green fluorescence) was used to stain <i>de novo</i> synthesized HaloTag α-actinin. Both red and green fluorescence were assessed simultaneously. Any newly synthesized α-actinin in different time intervals after TMRDirect staining would be labeled with R110Direct. Representative images were taken under identical exposure parameters. Slight variations in background fluorescence between durations could be due to cell-to-cell variation, differing locations on cover slips, slight variation in depth of mounting media, or to changes in focus.</p> 2015-06-15 02:50:46 intracellular proteases immunofluorescence studies sarcomere disassembly Sarcomere disassembly post-translational modifications actinin titin disassembly myocyte mitosis CDK 1-dependent cardiomyocyte mitosis Sarcomere dis sarcomeric Z-disk Cardiomyocyte Mitosis cyclin-dependent kinase 1 Dynamic Alterations cell cycle stages cells need heart muscle sarcomeric proteins cytosolic