Fig. 7 Inducible Hey2 depletion in adult mouse hearts enhances mitochondrial bioenergetics and cardiac function.a Design of induced Hey2 deletion by Myh6-MerCreMer. Tam, tamoxifen. b Western blots analysis of HEY2 in Hey2-iCKO and control hearts. 3 independent experiments. c Representative B-mode and M-mode echocardiography images of Hey2-iCKO and control hearts at 28 days post tamoxifen injection (dpi). d?f Analysis of LVEF, LVIDs and LVIDd of Hey2-iCKO and control hearts from 7 to 28 dpi. Control: n?=?15 (7 dpi,14 dpi), n?=?13 (21 dpi, 28 dpi). Hey2-iCKO: n?=?14 (7 dpi,14 dpi), n?=?12 (21 dpi, 28 dpi). Data presents as mean?±?SD, ****P?0.0001, Two-way ANOVA with Tukey multiple comparisons test. g RT-qPCR analysis of Ppargc1a, Esrra, Nrf1 and ETC complex genes in Hey2-iCKO and control hearts. 3 biological replicates. h RT-qPCR analysis of HEY2, PPARGC1 coactivator, ETC complex genes in DOX-treated and untreated AC16 cells. 3 biological replicates. i Diagram for tamoxifen and DOX injections, and hearts analysis in Hey2-iCKO and control mice. j, k Representative whole-mount hearts and quantification of heart weight (HW)/ body weight (BW) ratio from saline-injected control mice (Ctrl+saline, n?=?5), DOX-injected control mice (Ctrl+DOX, n?=?4), saline-injected Hey2-iCKO mice (iCKO+saline, n?=?5) and DOX-injected Hey2-iCKO mice (iCKO+DOX, n?=?3). Scale bar: 1000?µm. l, m Masson?s trichrome staining and fibrosis quantification of Ctrl+saline, Ctrl+DOX, iCKO+saline and iCKO+DOX hearts (n?=?8 mice). Scale bar: 100?µm. n Quantification of TUNEL+ cells in Ctrl+saline, Ctrl+DOX, iCKO+saline and iCKO+DOX hearts (n?=?6 mice). Box plots displayed median ± interquartile range (IQR), with whiskers extending to the minimum and maximum values. **P?0.01, ****P?0.0001, Student?s t-test (unpaired, two-tailed). o Analysis of LVEF, LVFS, LVIDs and LVIDd in hearts of Ctrl+saline (n?=?5), Ctrl+DOX (n?=?6), iCKO+saline (n?=?5) and iCKO+DOX (n?=?6) groups. p?r The serum levels of CK, CK-MB and LDH in Ctrl+saline (n?=?7), Ctrl+DOX (n?=?6), iCKO+saline (n?=?3) and iCKO+DOX (n?=?4) groups. Data presents as mean?±?SEM, *P?0.05, **P?0.01, ***P?0.001, ****P?0.0001, Student?s t-test (unpaired, two-tailed) (k, m, o?r) or Multiple t tests (two-tailed) (g, h). Source data are provided as a Source Data file.
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