Journal: Cell death & disease
Article Title: ELANE enhances KEAP1 protein stability and reduces NRF2-mediated ferroptosis inhibition in metabolic dysfunction-associated fatty liver disease.
doi: 10.1038/s41419-025-07603-2
Figure Lengend Snippet: Fig. 3 Elane regulates ferroptosis in hepatocytes via Gpx4. A Immunohistochemistry staining was used to assess Gpx4 expression in liver tissue after modelling in Elane+/+ and Elane−/−mice (n = 3). Scale bars: 100 μm. B, C Western blotting was used to assess Gpx4 protein expression in liver tissues after modelling in Elane+/+ and Elane−/−mice, and a graded analysis was performed (n = 3). D qRT‒PCR was used to assess the mRNA expression of Gpx4 in the liver tissues of Elane+/+ and Elane−/−mice after modelling (n = 3). E Extraction of protein from primary mouse hepatocytes, followed by Western blotting to assess Gpx4 protein levels after FFA and/or Elane treatment, after which grayscale analysis was performed (n = 3). F, G Western blotting was used to assess Gpx4 protein levels after the overexpression of Gpx4 by FFA and/or Elane treatment, after which grayscale analysis was performed (n = 3). H MDA levels after the overexpression of Gpx4 following FFA and/or Elane treatment (n = 3). I CCK-8 was used to assess cell viability after the overexpression of Gpx4 after FFA and/or Elane treatment (n = 3). J, K Extraction of protein from primary hepatocytes from Elane+/+ and Elane−/−mice after modelling, followed by Western blotting to assess Gpx4 protein expression, after which grayscale analysis was performed (n = 3). L MDA levels in primary hepatocytes from Elane+/+ and Elane−/−mice after modelling were assessed after 24 h of treatment with the Gpx4 inhibitor RSL3 (2 μM) (n = 3). M Cell viability of primary hepatocytes after modelling in Elane+/+ and Elane−/−mice was assayed by CCK-8 after the addition of RSL3 for 24 h (n = 3). The data are presented as the means ± SDs. *P < 0.05, **P < 0.01, in comparison to the control group; ns not significant.
Article Snippet: The levels of Elane in the tissue homogenate and plasma from different groups were quantified with a mouse Elane ELISA kit (E-EL-M3025, Elabscience) following the manufacturer’s instructions.
Techniques: Immunohistochemistry, Staining, Expressing, Western Blot, Extraction, Over Expression, CCK-8 Assay, Comparison, Control