Journal: Cells
Article Title: The Capability to Undergo ACSL4-Mediated Ferroptosis Is Acquired During Brown-like Adipogenesis and Affected by Hypoxia
doi: 10.3390/cells14161247
Figure Lengend Snippet: Effects of hypoxia in SGBS organoids. ( a – c ) Western blot analysis of normoxic (N) and hypoxic (H) SGBS organoids. n = 3; one-sample t test. ( d ) Analysis of protein oxidation by OxyBlot. A representative result of n = 3 independent experiments is shown. The DNPH substrate was added (+) or omitted (−) to confirm signal specificity. PVDF membrane was counterstained with EZBlue. ( e ) SA β-Gal staining of SGBS organoids. Grid: 100 µm. ( f ) Cytokine array. Cell culture supernatants of normoxic and hypoxic SGBS organoids of n = 3 experiments were pooled and subjected to cytokine measurement. TAT-Cyclo was used to inhibit HIF-1α. Top ten hypoxia-induced cytokines are indicated: (1) IL-6R, (2) CCL3, (3) TPO, (4) CCL27, (5) CCL16, (6) BTC, (7) CXCL8, (8) Adiponectin, (9) TIMP-2 and (10) TIMP-1. (+) Array positive control. Spearman correlation was used to analyze various parameters in clinical samples. p values ≤ 0.05 were considered to be significant as indicated: * = p < 0.05.
Article Snippet: Protein samples (5–10 μg) were separated by polyacrylamide gel electrophoresis and blotted onto a Polyvinyl Difluoride (PVDF) membrane with the semi-dry TransBlot Turbo Transfer System (BioRad).
Techniques: Western Blot, Membrane, Staining, Cell Culture, Positive Control