Journal: JCI Insight

Article Title: Cold stress–induced ferroptosis in liver sinusoidal endothelial cells determines liver transplant injury and outcomes

doi: 10.1172/jci.insight.174354

Figure Lengend Snippet: ( A ) Western blot–assisted detection and relative intensity ratio of NRF2, CHOP, MICU1, and MDA in cold-stressed LSECs (6 h/4°C). Expression of β-actin served as the internal control and was used for normalization ( n = 3/group). ( B ) Representative ( n = 6/group) images of dead cell detection in WT and NRF2-KO LSECs with/without MICU1 inhibitor (MCU-i4; 10 μM). Arrowheads indicate dead cells. Scale bars = 100 μm. ( C ) Quantification of dead cells ( n = 6/group). ( D ) Representative ( n = 3/group) immunohistochemical staining of MDA/NRF2 in WT and NRF2-KO LSECs with/without MICU1 inhibitor (MCU-i4). Scale bars = 100 μm. Arrows indicate nuclear localization of NRF2. Purple circle: WT; green circle: WT+MCU-i4; pink circle: NRF2-KO; yellow circle: NRF2-KO+MCU-i4. Data are shown as mean ± SEM. * P < 0.05, ** P < 0.01, Student’s t test ( A and D ).

Article Snippet: Fer-1 (SML0583, MilliporeSigma) and MCU-i4 (S9842, Selleck Chemicals) were used at 10 μM according to previous reports ( , ).

Techniques: Western Blot, Expressing, Immunohistochemical staining, Staining

Journal: JCI Insight

Article Title: Cold stress–induced ferroptosis in liver sinusoidal endothelial cells determines liver transplant injury and outcomes

doi: 10.1172/jci.insight.174354

Figure Lengend Snippet: ( A ) WT and NRF2- KO livers stored in UW solution (4°C/18 h) with/without MICU1 inhibitor (MCU-i4; 10 μM) were perfused with PBS (2 mL) through a cuff placed at the portal vein to collect liver flush from inferior vena cava. They were then transplanted into WT recipients ( n = 5−6/group). Serum and OLT samples were analyzed at 6 hours after reperfusion. The sham group underwent the same procedures except for OLT. ( B ) Western blot–assisted detection of MDA and HMGB1 levels in the liver flush (5 μL) from cold-stored livers ( n = 3−4/group). ( C ) LDH levels (U/L) in the liver flush ( n = 4−6/group). ( D ) Representative ( n = 5−6/group) H&E and TUNEL staining. Scale bars = 100 μm. ( E ) Suzuki’s histological grading of liver IRI and quantification of TUNEL-positive cells/HPF in OLT. ( F ) Serum AST and ALT levels (U/L). ( G ) Representative ( n = 5−6/group) integrin αIIb staining. Scale bars = 100 μm. ( H ) Western blot–assisted detection and relative intensity ratio of integrin αIIb in OLT. Expression of β-actin served as the internal control and was used for normalization ( n = 4/group). White circle: WT sham; black circle: NRF2-KO sham; purple circle: WT OLT; pink circle: NRF2-KO OLT; yellow circle: NRF2-KO+MCU-i4 OLT. Data are shown as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, 1-way ANOVA followed by Dunnett’s multiple comparisons test ( B , C , E , F , and H ).

Article Snippet: Fer-1 (SML0583, MilliporeSigma) and MCU-i4 (S9842, Selleck Chemicals) were used at 10 μM according to previous reports ( , ).

Techniques: Western Blot, TUNEL Assay, Staining, Expressing

Journal: Advanced Science

Article Title: Gastrointestinal Incomplete Degradation Exacerbates Neurotoxic Effects of PLA Microplastics via Oligomer Nanoplastics Formation

doi: 10.1002/advs.202401009

Figure Lengend Snippet: Stabilizing mitochondrial calcium levels ameliorates PLA MP‐induced PD‐like neurodegeneration in mice. A) Treatment strategy and experimental design overview. B) Relative mitochondrial calcium concentration and C) relative ATP content in midbrains after consecutive 28 days of exposure to PLA polymer MPs while treated with MCU‐i4 or DBcAMP. D) Activity levels in the open field test. E) Grip strength assessed in the grip strength test. F) Latency time measured in the rotarod test. G) Representative images of Nissl staining, TH staining, and TUNEL staining in mouse midbrains. Relative percentages of positive staining in midbrain cells for H) Nissl staining, I) TH staining, and J) TUNEL staining. Statistical analyses are determined by ANOVA, followed by Tukey's multiple comparison tests; * p < 0.05. ANOVA, analysis of variance; Micu3 , mitochondrial calcium uptake family member 3; MPs, microplastics; PLA, polylactic acid. TH, tyrosine hydroxylase; TUNEL, TdT‐mediated dUTP nick‐end labeling.

Article Snippet: MCU‐i4 (MCU inhibitor; Glpbio, China) or DBcAMP (NCLX agonist; Glpbio, China) and other chemicals were sourced from commercial suppliers with the highest available purity.

Techniques: Concentration Assay, Polymer, Activity Assay, Staining, TUNEL Assay, Comparison, End Labeling