Journal: Molecular Medicine Reports

Article Title: Reoxygenation induces reactive oxygen species production and ferroptosis in renal tubular epithelial cells by activating aryl hydrocarbon receptor

doi: 10.3892/mmr.2020.11679

Figure Lengend Snippet: AhR activation status does not affect Nrf2 activation or transcriptional activity. RPTECs were cultured under ctrl conditions or subjected to Reox with or without the AhR inhibitor CH223191. (A) Representative western blots of Nrf2 levels (corresponding to its activation status) and the expression of the Nrf2 transcriptional targets xCT (SLC7A11) and SOD-3. (B-D) Statistical analysis of the western blots. Neither Reox nor CH223191 affects Nrf2 activity, or the expression of xCT and SOD-3. Data are presented as the mean ± SEM of six independent experiments. AhR, arylhydrocarbon receptor; Nrf2, nuclear factor erythroid 2-related factor 2; xCT, cystine-glutamate antiporter; SOD-3, superoxide dismutase; RPTEC, renal proximal tubular epithelial cell; ctrl, control; Reox, reoxygenation.

Article Snippet: Primary antibodies were specific for AhR (1:200; cat. no. sc-133088; Santa Cruz Biotechnology, Inc.), cytochrome P450 family 1 subfamily A member 1 (CYP1A1; 1:500; cat. no. sc-25304; Santa Cruz Biotechnology, Inc.), Nrf2 (1:1,000; cat. no. TA343586; OriGene Technologies, Inc.), superoxide dismutase 3 (SOD-3; 1:100; cat. no. sc-271170; Santa Cruz Biotechnology, Inc.), cystine-glutamate antiporter (xCT, also known as SLC7A11; 1:1,000; cat. no. ANT-111; Alomone Labs), HIF-1α (1:500; cat. no. sc-10790; Santa Cruz Biotechnology, Inc.), LDH-A (1:1,000; cat. no. 2012; Cell Signaling Technology, Inc.), activated cleaved caspase-3 (CC3; 1:1,000; cat. no. ab13847; Abcam) and β-actin (1:2,500; cat. no. 4967; Cell Signaling Technology, Inc.).

Techniques: Activation Assay, Activity Assay, Cell Culture, Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: Loss of STARD7 Impairs Mitochondrial Phospholipid Homeostasis and Contributes to Mitochondrial Myopathy

doi: 10.1101/2025.11.27.691062

Figure Lengend Snippet: Loss of STARD7 disrupts fast-twitch–associated gene expression and fiber-type composition in soleus muscle (A) PCA of RNA-seq data from control (cont) and mKO soleus muscles. Each dot represents an individual animal (n = 3 per group). (B) Volcano plot of differentially expressed genes (DEGs). Differential expression was analyzed using the edgeR package after applying filterByExpr to remove low-expression genes. Genes with |log fold change| > 1 and FDR < 0.05 were defined as DEGs. Fast-twitch–related genes, including Mybpc2 , Actn3 , Pvalb , Mct4 , and Myh4 , were significantly downregulated in mKO. (C) Heatmap of DEGs (FDR < 0.05, |log fold change| > 1) in the soleus. (D) Functional enrichment analysis of DEGs using the clusterProfiler package for Gene Ontology (GO) and WikiPathways. GO terms and pathways with adjusted p < 0.05 were considered significantly enriched. (E) Immunoblot analysis of fast-twitch markers. Each lane represents an independent animal (n = 3). CBB staining confirms equal loading. (F) Representative image of soleus cross-sections stained with isoform-specific myosin heavy chain antibodies. Blue, type I (MyHC-I); red, type IIa (MyHC-IIa); green, type IIb (MyHC-IIb). Fibers lacking these isoforms were classified as type IIx. Scale bar, 200 μm.

Article Snippet: Antibodies used were anti-STARD7 (15191-1-AP), UQCRC2 (14742-1-AP), PVALB (29312-1-AP), MYBPC2 (25257-1-AP), MCT4 (22787-1-AP), MYH4 (20140-1-AP), MYH7 (22280-1-AP), CRLS1 (14845-1-AP), and PGS1 (17149-1-AP) (Proteintech, Rosemont, IL, USA); anti-TAZ (sc-365810) (Santa Cruz Biotechnology, Dallas, TX, USA); OXPHOS complexes (ab110413) (Abcam, Cambridge, UK); anti-OPA1 (612606) (BD Biosciences, San Jose, CA, USA); and 4-HNE (MHN-100P) (JaICA, Shizuoka, Japan).

Techniques: Gene Expression, RNA Sequencing, Control, Muscles, Quantitative Proteomics, Expressing, Functional Assay, Western Blot, Staining