Deletion of ATGL in adipose tissue attenuates pressure overload-induced LV failure.

<p><b>A:</b> Representative images of the hearts. <b>B</b>: Heart weight (HW)/ body weight (BW) ratio (mean and SEM, n = 5–6). <b>C</b>: Representative microscopic cross-sections of the hearts stained with hematoxylin/ eosin (H/E). <b>D</b>: Myocardial area, calculated based on microscopic sections of heart tissue, stained with H/E, analogue to the images presented in C (mean and SEM, n = 5–6). <b>E</b>: Representative M-Mode images of the echocardiographic analysis. <b>F-J</b>: cardiac echocardiographic analysis of mice (mean and SEM, n = 7): <b>F</b>: Left-ventricular mass (LVM). <b>G</b>: LVM relative to tibia length (LVM/TL). <b>H</b>: Left-ventricular internal diameter in diastole (LVID-d). <b>I</b>: Ejection fraction [%] (EF). <b>J</b>: Fractional shortening [%] (FS). <b>K</b>: Analysis of mRNA expression of beta-cardiac myosin heavy chain isogene (βMHCH), qRT-PCR studies were carried out using total RNA isolated from LV tissue. Data are presented as x-fold over wt-sham mice (mean and SEM, n = 5–6). <b>L</b>: Representative microscopic cross-sections of the hearts stained with picrosirius red. <b>M</b>: Representative high magnification images from picrosirius red-stained sections. <b>N:</b> Cardiac fibrosis calculated based on microscopic sections of the heart tissue, stained with picrosirius red, analogue to the images presented in L: 0 = no fibrosis, 1 = mild fibrosis, 2 = moderate fibrosis, 3 = severe fibrosis (mean and SEM, n = 5–6). <b>O and P:</b> Analysis of mRNA expression of collagen (Col) 1a1 (O) and Col3 (P), qRT-PCR studies were carried out using total RNA isolated from LV tissue. Data are presented as x-fold over wt-sham mice (mean and SEM, n = 5–6).*p<0.05 vs. wt sham, **p<0.01 vs. wt sham, ***p<0.001 vs. wt sham, ****p<0.0001 vs. wt sham, $p<0.05 vs. wt TAC,$ $p<0.01 vs. wt TAC,$ $$ p<0.001 vs. wt TAC, $$ $$ p<0.0001 vs. wt TAC, ##p<0.01 vs. atATGL-KO sham; 2-way ANOVA (Bonferroni post-test).</p

Selected lipid species are altered in plasma samples from patients with HFrEF MS-based shotgun lipidomics analysis of human plasma samples from HFrEF-patients (n = 13) and non-HFrEF controls (n = 10).

<p>A: Box plots show distribution of total mole percent values for each lipid class corrected for age and BMI (see supplementary methods). To test for differential changes a Mann-Whitney U test between HFrEF-patients and controls was performed. Adjusted p-values are indicated: *p<0.05, **p<0.01, **p<0.001. B: Estimated mean log2 fold change (HFrEF vs. control) vs. estimated mean mole percent of lipid species in the control group (see supplementary methods). Triangles represent significantly changed lipid species (FDR adjusted p-value < 0.1 and absolute value of log2-fold change ≥ 0.5), bubbles show those which are not significantly changed, size indicates log-transformed adjusted p-values. C: Bar graph shows the estimated log2-fold change (HFrEF vs. control) ± regression standard error of differentially changed lipid species (see B.). Colors represent log10 adjusted p-values as indicated. Lipid classes: Cer: ceramide, DAG: diacylglycerol, LPC: lyso-phosphatidylcholine, LPE: lyso-phosphatidylethanolamine, PC: phosphatidylcholine, PC O-: phosphatidylcholine-ether, PE: phosphatidylethanolamine, PE O-: phosphatidylethanolamine-ether, PI: phosphatidylinositol, SE: sterol ester, SM: sphingomyelin, ST: sterols, TAG: triacylglycerol.</p

Induction of pressure-mediated cardiac PE species is attenuated in atATGL-KO mice.

<p>MS-based shotgun lipidomics analysis of heart tissue samples (LV) isolated 11 weeks after intervention (sham or TAC) from wild-type (wt) or atATGL-KO mice. Lipid class denotations see <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1007171#pgen.1007171.g003" target="_blank">Fig 3</a>. <b>A-F:</b> Mean log2-fold change (TAC vs. sham) vs. mean mole percent of lipid species. Triangles represent significantly changed lipid species (FDR adjusted p-value < 0.1 and absolute value of log2-fold change ≥ 0.5); bubbles represent those which are not significantly changed; size indicates log-transformed adjusted p-values. <b>A, C, E:</b> wt-mice. <b>B, D, F:</b> atATGL-KO-mice. <b>G+H:</b> Significantly changed PC-PE ratios of matched FAs in wt-mice (<b>G</b>) or atATGL-KO mice (<b>H</b>). The mean ratio ± SEM is shown on a logarithmic scale, Mann-Whitney U test for TAC vs. sham: *p<0.05, **p<0.01 (adjusted) in wt-mice, no significant changes were found in at ATGL-KO mice. <b>I:</b> upper panels: WB analysis of heart lysates using antibodies against cleaved caspase 3 and Bcl-associated X protein (Bax); lower panel: WB analysis of HL-1 cardiomyocytes lysates from cells stimulated with vehicle (Veh) or fatty acid (FA) mix (C16:0, C18:1,C18:2 in different concentrations) using antibodies against cleaved caspase 3; loading control: β–actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH).</p

Metabolic phenotype and analysis of blood FA-profile in wt- and atATGL-KO mice.

<p><b>A:</b> Intraperitoneal Glucose Tolerance Test (ipGTT), (n = 4–6, 2-way ANOVA (Bonferroni post-test) from AUC). <b>B</b>: Area under the curve (AUC) of ipGTT, (mean and SEM, n = 4–6, 2-way ANOVA (Bonferroni posttest)). <b>C</b>: Insulin Tolerance Test (ITT), (n = 7–8, 2-way ANOVA (Bonferroni posttest) from AUC). <b>D</b>: Area under the curve (AUC) of ITT, (mean and SEM, n = 7–8, 2-way ANOVA (Bonferroni posttest)). <b>E</b>: Profile of selected serum FAs in TAC-operated mice analyzed by rapid resolution HPLC/ Tandem MS. <b>F</b>: Serum level of non-esterified fatty acids (NEFAs) in wt-TAC and atATGL-KO-TAC mice. <b>G</b>: Serum level of triacylglyerols (TAGs) in wt-TAC and atATGL-KO-TAC mice. (mean and SEM, n = 5, or as indicated, unpaired t-test). ***p<0.001 vs. wt sham, $p<0.05 vs. wt TAC,$ $$ p<0.001 vs. wt TAC, $$ $$ p<0.001 vs. wt TAC.</p