Fig 1: Inhibition of BECN1 expression by NaHS attenuates ferroptosis in cardiomyocytes. (A) Western blotting of BECN1 protein expression levels in BECN1-knockdown cells. (B) Viability of BECN1-knockdown cells following induction with 5 µg/ml LPS. (C) Western blotting of ferroptosis- and BECN1/SLC7A11 signaling-associated protein expression levels in H9c2 cells. *P<0.05 vs. LPS; #P<0.05 vs. LPS + NaHS. LPS, lipopolysaccharide; NaHS, sodium hydrosulfide; BECN1, Beclin 1; si, small interfering; NC, negative control; SLC7A11, solute carrier family 7 member 11; GPX4, glutathione peroxidase 4.
Fig 2: NaHS inhibits myocardial ferroptosis and improves sepsis-induced cardiomyopathy. (A) Cardiac ultrasound and cardiac function index. (B) Representative images of H&E staining of rat myocardial sections. (C) Representative images of mitochondrial transmission electron microscopy of rat myocardium. (D) Western blotting of ferroptosis- and BECN1/SLC7A11 signaling-associated protein expression levels in rat myocardial tissue. *P<0.05 vs. Sham; #P<0.05 vs. CLP. H&E, hematoxylin and eosin; CLP, cecal ligation and puncture; NaHS, sodium hydrosulfide; BECN1, Beclin 1; SLC7A11, solute carrier family 7 member 11; GPX4, glutathione peroxidase 4; p-, phosphorylated.
Fig 3: NaHS inhibits sepsis-induced ferroptosis in cardiomyocytes. (A) Ferroptosis regulatory protein expression levels and (B) fluorescence co-localization of BECN1 and SLC7A11 in H9c2 cells treated with 5 µg/ml LPS and 50 µmol/l NaHS. *P<0.05 vs. control; #P<0.05 vs. LPS. LPS, lipopolysaccharide; NaHS, sodium hydrosulfide; BECN1, Beclin 1; SLC7A11, solute carrier family 7 member 11; PCC, Pearson's correlation coefficient; MCC, Mander's co-localization coefficient; GPX4, glutathione peroxidase 4; p-, phosphorylated.
Fig 4: The mechanism of miR-17-5p is related to the Akt/Beclin1 signaling pathway. Representative band and statistical analysis (A) of p-Akt, Akt, Beclin1, p-Beclin1(234), and p-Beclin1 (295) expression among HK-2 cells treated with hypoxia or the miR-17-5p agomir. Representative band and statistical analysis (B) of p-Akt, Akt, Beclin1, p-Beclin1 (234), and p-Beclin1 (295) expression among HK-2 cells, which were treated with hypoxia, pcDNA3.1-PTEN, pcDNA3.1-BIM, or related siRNAs. Western blot (C) confirmed that the miR-17-5p agomir promoted the expression of p-AKT and Akt, while the Akt-siRNA reversed the upregulation. Akt-siRNA significantly increased the LC3II/I ratio and decreased the expression of p62, while the miR-17-5p agomir reversed this effect (D). Flow cytometry suggested that Akt-siRNA greatly increased the apoptosis of HK-2 cells. *, P<0.05. ago-NC, agomir-negative control; Akt, protein kinase B; BIM, Bcl-2 like protein 11; GAPDH, glyceraldehyde phosphate dehydrogenase; HK-2, human kidney-2; LC3, microtubule-associated protein light chain 3; PTEN, phosphatase and tensin homolog deleted on chromosome 10; siRNA, small interfering RNA.
Fig 5: The miR-17-5p agomir was administrated via the tail vein 24 h before I/R modeling (n=4 per group). The expression of miR-17-5p in mice kidneys (A), and the concentration of BUN and Scr (B) in mice after 24 h and 48 h of intervention. Representative images of kidney histology (C) with H&E and TUNEL staining, and the statistical analysis of apoptosis in kidneys. Representative band and statistical analysis (D) of LC3B II/I and P62 expression in mice kidneys. Representative images of autophagosome and autophagolysosome (E) in mice kidneys detected by TEM, and the red arrow stands for autophagosome and autophagolysosome. The mRNA expression of BIM and PTEN in mice kidneys were detected by real-time PCR (F). Representative band and statistical analysis (G) of BIM, PTEN, p-Akt, Akt, Beclin1, p-Beclin1 (234), and p-Beclin1 (295) expression in mice kidneys. Trends in the concentrations of BUN and Scr (H) in mice for 7 days postoperative follow-up. *, P<0.05. ago-NC, agomir-negative control; Akt, protein kinase B; BIM, Bcl-2 like protein 11; BUN, blood urea nitrogen; DAPI, 4’,6-diamidino-2-phenylindole; GAPDH, glyceraldehyde phosphate dehydrogenase; H&E, hematoxylin-eosin; I/R, ischemia and reperfusion; LC3, microtubule-associated protein light chain 3; PTEN, phosphatase and tensin homolog deleted on chromosome 10; Scr, serum creatinine; TEM, transmission electron microscopy; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling.
Supplier Page from Abcam for Anti-Beclin 1 (phospho S295) antibody