Journal: Science Advances

Article Title: The mechanism of action of digoxin requires the sodium-dependent inactivation of the sodium-calcium exchanger

doi: 10.1126/sciadv.ady9596

Figure Lengend Snippet: ( A ) Representative Western blot of NKA α-subunit (top) and GAPDH (bottom) expression of WT and K229Q ventricular homogenates. ( B ) Summary densitometry analysis showing the fold change in NKA protein expression between WT and K229Q mice, represented as NKA intensity/GAPDH intensity, normalized to WT levels. Hearts: WT, n = 10; K229Q, n = 10. Immunofluorescence image of WT ( C ) and K229Q ( D ) adult ventricular myocytes. Cells were stained with anti-NKA α-subunit antibody. The panel shows the magnified area of the cell. Cells in the image are maximum intensity projections. Scale bar, 10 μm. ( E ) NKA fluorescence is reported as corrected total cell fluorescence (CTCF) divided by cell area (CTCF/area). Each point represents one animal. Cells (four to six) were averaged for each animal. Animal averages were used for statistical comparisons. ( F ) Data from each individual cell, as in (E). NKA fluorescence was unchanged in K229Q myocytes compared to WT. Data are means ± SEM. Cells/animals: WT, n = 15/3; K229Q, n = 15/3.

Article Snippet: Myocytes were then incubated with the mouse monoclonal anti-NKA α-subunit antibody (1:200, a5, DSHB) in 3% BSA PBS overnight at 4°C.

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Fluorescence

Journal: Science Advances

Article Title: The mechanism of action of digoxin requires the sodium-dependent inactivation of the sodium-calcium exchanger

doi: 10.1126/sciadv.ady9596

Figure Lengend Snippet: ( A ) Representative Western blot of NKA α-subunit (top) and GAPDH (bottom) expression of WT and K229Q ventricular homogenates. ( B ) Summary densitometry analysis showing the fold change in NKA protein expression between WT and K229Q mice, represented as NKA intensity/GAPDH intensity, normalized to WT levels. Hearts: WT, n = 10; K229Q, n = 10. Immunofluorescence image of WT ( C ) and K229Q ( D ) adult ventricular myocytes. Cells were stained with anti-NKA α-subunit antibody. The panel shows the magnified area of the cell. Cells in the image are maximum intensity projections. Scale bar, 10 μm. ( E ) NKA fluorescence is reported as corrected total cell fluorescence (CTCF) divided by cell area (CTCF/area). Each point represents one animal. Cells (four to six) were averaged for each animal. Animal averages were used for statistical comparisons. ( F ) Data from each individual cell, as in (E). NKA fluorescence was unchanged in K229Q myocytes compared to WT. Data are means ± SEM. Cells/animals: WT, n = 15/3; K229Q, n = 15/3.

Article Snippet: The blot was divided to probe for all α-subunit isoforms of the NKA (a5, Developmental Studies Hybridoma Bank) and GAPDH as a loading control.

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Fluorescence