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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and <t>QKI7</t> expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.
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Inducible QKI5, QKI6, and QKI7 expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.

Journal: Nature Communications

Article Title: QKI is a critical pre-mRNA alternative splicing regulator of cardiac myofibrillogenesis and contractile function

doi: 10.1038/s41467-020-20327-5

Figure Lengend Snippet: Inducible QKI5, QKI6, and QKI7 expression cells are established in H1-8 lines (H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind ). Representative images of immunofluorescence staining of TNNT2 and ACTN2 in H1-8- QKI5 ind , H1-8- QKI6 ind , H1-8- QKI7 ind cardiomyocytes with or without doxycycline induction. Myofibrillar structures are recovered in doxycycline-induced H1-8- QKI5 ind cardiomyocytes (Day-15) and ACTN2 expression is also recovered. However, doxycycline-induced H1-8- QKI6 ind and H1-8- QKI7 ind cardiomyocytes fail to recover ACTN2 expression and generate normal myofibrillar structures (Day-15). Scale bar: 25 μm. All experiments are independently repeated three times with three different sets of cell samples to confirm the reproducibility of the findings.

Article Snippet: The membranes were blocked with 5% nonfat dry milk (Bio-Rad, 1706404) in Tris-buffered saline containing Tween-20 and incubated with primary antibodies against TNNT2 (DSHB, Cat#AB528495), ACTN2 (Sigma, Cat#A7811), MF20 (DSHB, Cat#AB2147781), MYOZ2 (ThermoFisher, Cat#PA5-76946), TNNI3 (Santa Cruz, Cat#SC15368), Tubulin (Sigma, Cat#T6199), Pan-QKI(Abcam, Cat#ab126742), QKI-5 (Bethyl, A300-183A), QKI6 (Millipore, Cat#AB9906), and QKI7 (Millipore, Cat#AB9908), respectively, at 1 : 1000 dilution, and corresponding secondary antibodies labeled with horseradish peroxidase (Goat anti-Mouse IgG antibody, ThermoFisher Scientific, G21040; Mouse anti-Rabbit IgG antibody: Santa Cruz, sc-2357) at 1 : 4000 dilution.

Techniques: Expressing, Immunofluorescence, Staining