Figure 3.
Figure 3. Titin-eGFP dynamics are independent from protein synthesis. Cells were imaged for 14 h after photobleaching to follow recovery and treated with cycloheximide (CX) to distinguish titin mobility and de novo synthesis. (A) Images of control and CX-treated embryonic cardiomyocytes (E13.5) from homozygous titin-eGFP knockin mice were obtained before and immediately after bleaching (white arrows). Titin-eGFP was homogenously distributed in embryonic cardiomyocytes and fluorescence had significantly recovered after 14 h. Bar, 10 µm. (B) Plot of fluorescence intensity vs. time after photobleaching to compare titin-eGFP protein recovery of control and CX-treated cells. Inhibition of protein synthesis did not influence the mobility of titin-eGFP (control, n = 4; CX, n = 3). (C) Quantification of the titin-eGFP mobile fractions indicated a level of ∼50% in both control and CX-treated cardiomyocytes (control, n = 4; CX, n = 3). Error bars indicate SEM.

Titin-eGFP dynamics are independent from protein synthesis. Cells were imaged for 14 h after photobleaching to follow recovery and treated with cycloheximide (CX) to distinguish titin mobility and de novo synthesis. (A) Images of control and CX-treated embryonic cardiomyocytes (E13.5) from homozygous titin-eGFP knockin mice were obtained before and immediately after bleaching (white arrows). Titin-eGFP was homogenously distributed in embryonic cardiomyocytes and fluorescence had significantly recovered after 14 h. Bar, 10 µm. (B) Plot of fluorescence intensity vs. time after photobleaching to compare titin-eGFP protein recovery of control and CX-treated cells. Inhibition of protein synthesis did not influence the mobility of titin-eGFP (control, n = 4; CX, n = 3). (C) Quantification of the titin-eGFP mobile fractions indicated a level of ∼50% in both control and CX-treated cardiomyocytes (control, n = 4; CX, n = 3). Error bars indicate SEM.

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