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. Author manuscript; available in PMC: 2021 Mar 17.
Published in final edited form as: Circulation. 2020 Jan 29;141(11):916–930. doi: 10.1161/CIRCULATIONAHA.119.041882

Figure 1. Characterizing cell death dynamics and sphingolipid metabolism in mouse hearts after MI.

Figure 1.

A, Hearts were harvested from sham-operated mice or 1, 2, 4 and 28 days post MI. B, TUNEL assays were performed to assess DNA fragmentation in hearts harvested from either sham-operated mice or 1, 2, 4 or 28 days post MI. Troponin-I immunostaining was used to distinguish between cardiomyocytes and non-cardiomyocytes. Percentage of dead cells in left ventricle at day 1, 2, 4 and 28 post MI was quantified within all cardiac cells (DAPI+ cells) n=3 (C); CMs only (DAPI+, Troponin I+) n=3 (D) and non-CM cells only (DAPI+, Troponin I-) n=3 (E). F, For RNASeq, protein analysis and mass spectrometry, hearts were harvested from sham-operated mice or 4 or 24 hours post MI. RNA, proteins and lipids were extracted for sphingolipid determination. G, Hierarchical clustering dendrogram for the sphingolipid signaling pathway transcriptome in sham-operated hearts and hearts harvested 4 or 24 hours post MI, n=3, 3, 4, respectively. H, Sphingolipid levels measured in the LV of sham-operated mice or 24 hours post MI, n=3. I, Western blot of the AC precursor, AC active subunit β and Sphk1 in the LV of sham-operated hearts and hearts harvested 4 or 24 hours post MI. J, Quantified protein levels of AC precursor, AC β subunit and Sphk1, n=4. K, HPLC-MS/MS determination of AC activity in the LV of sham-operated hearts and hearts harvested 4 or 24 hours post MI, n=3. *, P<0.05, One-way ANOVA, Tukey’s Multiple Comparison Test for (J&K) and Holm-Sidak correction for multiple comparisons (H). Scale bar = 50μm. The results include two independent experiments for H-K.