GAS-STING signaling plays an essential pathogenetic role in Doxorubicin-Induced Cardiotoxicity

Figures

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  Fig. 1. The expression levels of cGAS and STING in the myocardium of DIC mice (A–E). A Echocardiographic images of control mice or DIC mice at day14. B left ventricle ejection fraction, left ventricle fractional shortening, left ventricular end-diastolic dimension, left ventricular end-systolic dimension at day 14 (n = 4). C-E 15 mg/Kg doxorubicin injection enhanced cGAS and STING expression in DIC mice (day 14) by immunoblot and mRNA analysis

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  Fig. 2. AAV9-sh-STING injection decreased heart STING expression levels of baseline conditions in vivo (A-E). A schematic representation of the experimental design. B representative images of fluorescence microscope of frozen heart left ventricular sections after 3 weeks of AAV9-control and AAV9-sh-STING injection (scale bar, 50 m). C-E immunoblot and mRNA analysis reveals AAV9-sh-STING injection reduced STING expression compared with AAV9-control injection after 3 week of injection

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  Fig. 3. Knockdown of STING improved survival and cardiac function of DOX-treated mice (A-E). A Effect of STING knockdown on the 14-day survival rate after DOX injection (n = 10). B Effect of STING knockdown on HW/TW. C Representative echocardiographic images of each group (n = 4). D Effect of STING knockdown on left ventricle ejection fraction, left ventricle fractional shortening, left ventricular end-diastolic dimension and left ventricular end-systolic dimension of each group (n = 4). E Effect of STING knockdown on DOX-induced LDH, cTNT and CK-MB release (n = 4)

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  Fig. 4. Light micrograph demonstrating the effect of STING knockdown on DOX-induced myocardial histological alterations. A Representative photomicrographs of mouse heart stained with H&E; B Representative photomicrographs of mouse heart stained with Masson;. Scale bar = 50 𝜇m. C Quantitative analysis for cytoplasmic vacuolization. D Quantitative analysis for myofibril loss. E Quantitative analysis for intestinal fibrosis (n = 4)

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  Fig. 5. STING knockdown suppressed cardiac apoptosis inflammation, morphological alteration of DIC mice (A-C). AThe protein levels of C-Caspase3, BAX and BCL-2 of each group after PBS or DOX injection. B TUNEL staining of each group after PBS or DOX injection (n = 4). The arrow indicated tunel positive cells. C mRNA analysis of inflammation-related genes of each group

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  Fig. 6. Suppressed DOX-induced phosphorylation of IRF3, TBK1 and p65 was observed by immunoblot analysis following STING knockdown in HL-1 cells following DOX treatment (1 μm for 24 h). Quantitative data represent the relative ratio to total IRF3, TBK1 or p65 (n = 6)

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  Fig. 7. cGAS-STING activation is involved in DOX-induced cell death in cultured cardiomyocytes (A-D). A siRNA-mediated STING knockdown influenced the protein levels of C-Caspase3, Caspase3, BAX and BCL-2 in indicated groups in HL-1 cells (n = 6). B-C Flow cytometry assay and TUNEL staining showed that STING knockdown reduced cell apoptosis (n = 4). D CCK8 assay detection reveals that STING knockdown increased cell viability. E STING knockdown suppressed the protein levels of NLPR3

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  Fig. 8. Schematic illustration of an impairment mechanism by cGAS-STING activation in DOX-induced cardiotoxicity