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Doxorubicin injection in 129S2/SvPasCrl males does not enhance proteolytic cleavage of <t>ENaC's</t> γ subunit. (a) A schematic is shown depicting cleavage sites in ENaC's γ subunit and the antibody used for immunoblot. (b) Immunoblot of ENaC's γ subunit from deglycosylated whole kidney lysates. Each lane represents a kidney lysate from one mouse. (c) Quantification of full‐length γ subunit or cleavage products (normalized to stain‐free gel protein intensity). Full‐length γ subunit and furin‐cleaved C‐terminal fragment abundances increase in doxorubicin‐treated mice compared to untreated controls. Distally cleaved γ subunit protein abundance does not increase significantly. (d) Abundance of the furin‐cleaved or distally cleaved proteolytic fragments relative to the total (full‐length + furin‐cleaved + distally cleaved) γ subunit abundance is not enhanced by treatment with doxorubicin. (Ctrl, control mice untreated with doxorubicin; Doxo, doxorubicin‐treated mice. Numbers in pair‐wise comparisons represent p values, as determined using Student's t ‐test.)
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Doxorubicin injection in 129S2/SvPasCrl males does not enhance proteolytic cleavage of <t>ENaC's</t> γ subunit. (a) A schematic is shown depicting cleavage sites in ENaC's γ subunit and the antibody used for immunoblot. (b) Immunoblot of ENaC's γ subunit from deglycosylated whole kidney lysates. Each lane represents a kidney lysate from one mouse. (c) Quantification of full‐length γ subunit or cleavage products (normalized to stain‐free gel protein intensity). Full‐length γ subunit and furin‐cleaved C‐terminal fragment abundances increase in doxorubicin‐treated mice compared to untreated controls. Distally cleaved γ subunit protein abundance does not increase significantly. (d) Abundance of the furin‐cleaved or distally cleaved proteolytic fragments relative to the total (full‐length + furin‐cleaved + distally cleaved) γ subunit abundance is not enhanced by treatment with doxorubicin. (Ctrl, control mice untreated with doxorubicin; Doxo, doxorubicin‐treated mice. Numbers in pair‐wise comparisons represent p values, as determined using Student's t ‐test.)
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Doxorubicin injection in 129S2/SvPasCrl males does not enhance proteolytic cleavage of <t>ENaC's</t> γ subunit. (a) A schematic is shown depicting cleavage sites in ENaC's γ subunit and the antibody used for immunoblot. (b) Immunoblot of ENaC's γ subunit from deglycosylated whole kidney lysates. Each lane represents a kidney lysate from one mouse. (c) Quantification of full‐length γ subunit or cleavage products (normalized to stain‐free gel protein intensity). Full‐length γ subunit and furin‐cleaved C‐terminal fragment abundances increase in doxorubicin‐treated mice compared to untreated controls. Distally cleaved γ subunit protein abundance does not increase significantly. (d) Abundance of the furin‐cleaved or distally cleaved proteolytic fragments relative to the total (full‐length + furin‐cleaved + distally cleaved) γ subunit abundance is not enhanced by treatment with doxorubicin. (Ctrl, control mice untreated with doxorubicin; Doxo, doxorubicin‐treated mice. Numbers in pair‐wise comparisons represent p values, as determined using Student's t ‐test.)
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Doxorubicin injection in 129S2/SvPasCrl males does not enhance proteolytic cleavage of <t>ENaC's</t> γ subunit. (a) A schematic is shown depicting cleavage sites in ENaC's γ subunit and the antibody used for immunoblot. (b) Immunoblot of ENaC's γ subunit from deglycosylated whole kidney lysates. Each lane represents a kidney lysate from one mouse. (c) Quantification of full‐length γ subunit or cleavage products (normalized to stain‐free gel protein intensity). Full‐length γ subunit and furin‐cleaved C‐terminal fragment abundances increase in doxorubicin‐treated mice compared to untreated controls. Distally cleaved γ subunit protein abundance does not increase significantly. (d) Abundance of the furin‐cleaved or distally cleaved proteolytic fragments relative to the total (full‐length + furin‐cleaved + distally cleaved) γ subunit abundance is not enhanced by treatment with doxorubicin. (Ctrl, control mice untreated with doxorubicin; Doxo, doxorubicin‐treated mice. Numbers in pair‐wise comparisons represent p values, as determined using Student's t ‐test.)
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Doxorubicin injection in 129S2/SvPasCrl males does not enhance proteolytic cleavage of ENaC's γ subunit. (a) A schematic is shown depicting cleavage sites in ENaC's γ subunit and the antibody used for immunoblot. (b) Immunoblot of ENaC's γ subunit from deglycosylated whole kidney lysates. Each lane represents a kidney lysate from one mouse. (c) Quantification of full‐length γ subunit or cleavage products (normalized to stain‐free gel protein intensity). Full‐length γ subunit and furin‐cleaved C‐terminal fragment abundances increase in doxorubicin‐treated mice compared to untreated controls. Distally cleaved γ subunit protein abundance does not increase significantly. (d) Abundance of the furin‐cleaved or distally cleaved proteolytic fragments relative to the total (full‐length + furin‐cleaved + distally cleaved) γ subunit abundance is not enhanced by treatment with doxorubicin. (Ctrl, control mice untreated with doxorubicin; Doxo, doxorubicin‐treated mice. Numbers in pair‐wise comparisons represent p values, as determined using Student's t ‐test.)

Journal: Physiological Reports

Article Title: Resistance to doxorubicin‐induced proteinuria and proteolytic activation of ENaC in 129S2 / SvPas mice

doi: 10.14814/phy2.70667

Figure Lengend Snippet: Doxorubicin injection in 129S2/SvPasCrl males does not enhance proteolytic cleavage of ENaC's γ subunit. (a) A schematic is shown depicting cleavage sites in ENaC's γ subunit and the antibody used for immunoblot. (b) Immunoblot of ENaC's γ subunit from deglycosylated whole kidney lysates. Each lane represents a kidney lysate from one mouse. (c) Quantification of full‐length γ subunit or cleavage products (normalized to stain‐free gel protein intensity). Full‐length γ subunit and furin‐cleaved C‐terminal fragment abundances increase in doxorubicin‐treated mice compared to untreated controls. Distally cleaved γ subunit protein abundance does not increase significantly. (d) Abundance of the furin‐cleaved or distally cleaved proteolytic fragments relative to the total (full‐length + furin‐cleaved + distally cleaved) γ subunit abundance is not enhanced by treatment with doxorubicin. (Ctrl, control mice untreated with doxorubicin; Doxo, doxorubicin‐treated mice. Numbers in pair‐wise comparisons represent p values, as determined using Student's t ‐test.)

Article Snippet: Lysates were then subjected to SDS‐PAGE and immunoblot using an antibody directed against the ENaC γ subunit's C‐terminus (StressMarq, #SPC‐405) as previously described (Ray et al., ).

Techniques: Injection, Western Blot, Staining, Quantitative Proteomics, Control