Cfz-induced cardiotoxicity mechanism: acute Cfz administration induced cardiotoxicity primarily through inhibition of AMPKα-mediated autophagy and PI3K/Akt/eNOS axis. (A) Representative western blots of protocol 2. (B) Representative column graphs and densitometry analysis of protocol 2: PI3K p85(Tyr458)/p55(Tyr199)/tPI3K, tPI3K/β-tubulin, pAkt(Ser473)/tAkt, tAkt/β-tubulin, peNOS(Ser1177)/teNOS, t-eNOS/β-tubulin, pAMPKα(Thr172)/tAMPKα, tAMPKα/β-tubulin, pmTOR(Ser2448)/tmTOR, tmTOR/β-tubulin, pRaptor(Ser792)/tRaptor, tRaptor/β-tubulin, LC3-II/ β-tubulin. (C) Representative western blots of protocol 3. (D) Representative column graphs and densitometry analysis of protocol 3: PI3K p85(Tyr458)/p55(Tyr199)/tPI3K, tPI3K/β-actin, PTEN/β-actin, pAkt(Ser473)/tAkt, tAkt/β-tubulin, peNOS(Ser1177)/teNOS, t-eNOS/β-tubulin, iNOS/β-tubulin, pAMPKα(Thr172)/tAMPKα, tAMPKα/β-actin, pmTOR(Ser2448)/tmTOR, tmTOR/β-tubulin, pRaptor(Ser792)/tRaptor, tRaptor/β-tubulin, LC3-II/β-tubulin. *P < .05, *P < .01, ***P < .001 n = 4-9 per group. In each protocol, 1 representative western blot image for the loading controls (ie, β-tubulin and/or β-actin) is presented for ergonomic reasons. Not all proteins ran on the same sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Western blots show 3 representative animals from each experimental group, 1 in each lane. IOD, integral optical density.