MKP5 deficiency protects against pressure overload-induced cardiac hypertrophy and fibrosis
Data files
Mar 04, 2024 version files 33.70 MB
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12-Image5.jpg
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13-Image14.jpg
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14-Image12.jpg
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15-Image16.jpg
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16-Image16.jpg
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202004017_erk_-2.tif
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202004017_p-erk_-5.tif
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202004025_p-p38_-2.tif
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202004025_p38_-1.tif
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202004026_jnk_-1.tif
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202004026_p-jnk_-2.tif
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249-Image8.jpg
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274-Image8.jpg
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275-Image8.jpg
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278-Image17.jpg
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PreTAC_12wks_TAC_Echo_data.xlsx
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README.md
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Abstract
Cardiac pressure overload resulting from transaortic constriction (TAC) induces MKP-5 expression and facilitates cardiac hypertrophy and fibrosis. In this dataset, we show that MKP-5 deficiency attenuates cardiac fibrosis after TAC. In vitro, bone marrow-derived macrophages (BMDMs) from MKP-5 knockout mice exhibit enhanced phosphorylation of p38 MAPK, JNK, and Erk compared to BMDMs from wild-type mice in response to IL-4 stimulation.
Data for MKP5 project funded by AHA
https://doi.org/10.5061/dryad.vdncjsz2v
The data set contains:
(1) Western blots for bone marrow-derived macrophages (BMDMs) from wild type and MKP5-/- mice stimulated with IL-4 for 0, 15, 30, 60, 90 min and blotted for p38 MAPK total and p38 MAPK phosphorylated at Thr180/Tyr182, JNK total and JNK phosphorylated at Thr183/Tyr185, Erk total and Erk phosphorylated at Thr202/Tyr204.
File 202004025 p38-1.tif: p38 MAPK, total. Left to right in order of wild type 0, 15, 30, 50, 90 min of IL-4 stimulation, followed by MKP5-/- 0, 15, 30, 60, 90 min of IL-4 stimulation.
File 202004025 p-p38-2.tif: p38 MAPK phosphorylated at Thr180/Tyr182. Left to right in order of wild type 0, 15, 30, 50, 90 min of IL-4 stimulation, followed by MKP5-/- 0, 15, 30, 60, 90 min of IL-4 stimulation.
File 202004026 jnk-1.tif: JNK total. Left to right in order of wild type 0, 15, 30, 50, 90 min of IL-4 stimulation, followed by MKP5-/- 0, 15, 30, 60, 90 min of IL-4 stimulation.
File 202004026 p-jnk-2.tif: JNK phosphorylated at Thr183/Tyr185. Left to right in order of wild type 0, 15, 30, 50, 90 min of IL-4 stimulation, followed by MKP5-/- 0, 15, 30, 60, 90 min of IL-4 stimulation.
File 202004017 erk-2.tif: Erk total. Left to right in order of wild type 0, 15, 30, 50, 90 min of IL-4 stimulation, followed by MKP5-/- 0, 15, 30, 60, 90 min of IL-4 stimulation.
File 202004017 p-erk-5.tif: Erk phosphorylated at Thr202/Tyr204. Left to right in order of wild type 0, 15, 30, 50, 90 min of IL-4 stimulation, followed by MKP5-/- 0, 15, 30, 60, 90 min of IL-4 stimulation.
(2) Masson's Trichrome staining of mouse 12, 13, 14, 15, 16 (MKP5-/-) and 249, 274, 275, 278 (MKP5+/+) at 12 wks after TAC. Blue staining is fibrosis. The files are named as mouse ID-image#, e.g., 12-image5.jpg means image #5 for mouse 12.
(3) Echo cardiography analysis (File: PreTAC + 12wks TAC Echo data) of mouse 12, 13, 14, 15, 16 (genotype: MKP5-/-) and 249, 274, 275, 278 (genotype: MKP5+/+) at Pre-TAC and 12 weeks post TAC. Parameters measured include interventricular septum (IVS) thickness (mm) at end of diastole (d) (column D) and at end of systole (s) (column E), left ventricle internal diameter (mm) end distole (F) and end systole (G), left ventricle posterior wall thickness (mm) end distole (H) and end systole (I), ejection fraction (%, J), fractional shortening (%, K), left ventricular mass (mg) (L) and left ventricular mass (mg) corrected (M), left ventricle volume (ul) end distole (N) and end systole (O).
Wild-type and MKP-5 knockout mice were subjected to TAC surgery. Cardiac function was subsequently analyzed by Echo cardiography with a VisualSonics Vevo 2100 machine. Fixed mouse hearts were processed for paraffin sections and Masson's trichrome staining to visualize fibrosis. Fibrosis was quantified using Image J software.
Bone marrow-derived macrophages from wild type and MKP-5 knockout mice were stimulated with IL-4 in culture for 0, 15, 30, 60, and 90 min and then lysed in RIPA buffer to collect protein for Western blots. Protein was separated by 10-15% SDS-PAGE and blotted for p38 MAPK total and p38 MAPK phosphorylated at Thr180/Tyr182, JNK total and JNK phosphorylated at Thr183/Tyr185, Erk total and Erk phosphorylated at Thr202/Tyr204. Immunoblots were developed using ODYSSEY infrared Imaging SYstem (LI-COR) and quantified with Image Studio software (LI-COR version 5.2).