FIGURE

Fig. 4

ID

ZDB-FIG-210117-28

Publication

Kuo et al., 2021 - Yulink, predicted from evolutionary analysis, is involved in cardiac function

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Fig. 4

Over-expression of YULINK, PPARγ and SERCA2 rescued the phenotypes of mouse Yulink KD HL-1 cardiomyocytes. Control (Ctrl), YULINK, PPARγ or SERCA2 plasmids (carrying BFP as indicator of expression marker) were transfected or electroporated into Yulink KD cells, and then analyzed for Ca²⁺ cycling using fluorescent Ca²⁺ dye, Rhod-2 AM. a Over-expression of YULINK or PPARγ. Representative line-scan images and spontaneous Ca²⁺ transient in Yulink KD mouse HL-1 cardiomyocytes. Arrhythmia-like waveforms were observed in Yulink KD cardiomyocytes with Ctrl vector (red arrows). After over-expression of YULINK (YULINK-OE) or PPARγ (PPARγ-OE) in Yulink KD cells, normal waveforms were observed. The result of Ca²⁺ spark analysis was consistent with the increase of Ca²⁺ transient amplitudes (3.1 ± 0.5 for YULINK-OE or 3.2 ± 0.3 for PPARγ-OE, vs. 1.9 ± 0.35 for Ctrl). The cells with YULINK or PPARγ over-expression also exhibited a decrease of the percentages of irregular Ca²⁺ transients (8 ± 2% for YULINK-OE or 10 ± 2% for PPARγ-OE, vs. 27 ± 3% for Ctrl). The cells with YULINK or PPARγ over-expression also exhibited a reduction of the Ca²⁺ decay (Tau) rate (430 ± 42 ms for YULINK-OE or 450 ± 51 ms for PPARγ-OE, vs. 605 ± 47 ms for Ctrl). Ctrl (black bars), YULINK-OE (orange bars) and PPARγ-OE (blue bars) (n = 40, each). *p < 0.05, Student’s t test. b Over-expression of SERCA2. Representative line-scan images and spontaneous Ca²⁺ transients in Yulink KD mouse HL-1 cardiomyocytes. Arrhythmia-like waveforms were observed in Yulink KD cardiomyocytes with control vector (Ctrl, red arrows). After over-expression of SERCA2 (SERCA2-OE) in Yulink KD cells, normal waveforms were observed. The result of Ca²⁺ spark analysis was consistent with the increase of Ca²⁺ transient amplitudes (3.2 ± 0.45 for SERCA2-OE vs. 2.02 ± 0.3 for Ctrl). The cells with SERCA2-OE also exhibited a decrease of the percentages of irregular Ca²⁺ transients (11 ± 3% for SERCA2-OE vs. 32 ± 5% for Ctrl). The cells with SERCA2-OE also exhibited a reduction of the Ca²⁺ decay (Tau) rate (412 ± 50 ms for SERCA2-OE vs. 635 ± 78 ms for Ctrl). Ctrl vector (black bars), SERCA2-OE (gray bars) (n = 35, each). *p < 0.05, Student’s t test

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ J. Biomed. Sci.