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