Journal: The Journal of Clinical Investigation

Article Title: Congenital deficiency reveals critical role of ISG15 in skin homeostasis

doi: 10.1172/JCI141573

Figure Lengend Snippet: Human iPSCs carrying the WT ISG15 locus or a CRISPR/Cas9-mediated targeted deletion were differentiated into CD31-positive vascular ECs and stimulated with IFN-α (1000 IU/mL) for 12 or 24 hours. ( A ) Absence of ISG15 mRNA in ISG15 –/– ECs. ( B – E ) Increase in intracellular IFIT2 mRNA, and IP10 and IL-1β but not TNF-α protein in culture supernatants. ( F – L ) Decreased expression in several collagen mRNAs but not COL1A1 , MMP1 , and TIMP1 in ISG15 –/– ECs. RT-qPCR with ΔΔCt analysis; reference = unstimulated WT cells ( A , B , and F – L ), or enzyme-linked immunoassay ( C – E ). n = 3, ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001 (Student’s t test).

Article Snippet: On day 7 of differentiation, CD31-positive cells were purified by Magnetic Activated Cell Sorting using CD31 MicroBead Kit (Miltenyi Biotec). hiPSC-ECs were then cultured in ECGM-2 medium (PromoCell) on plates coated with fibronectin (Corning).

Techniques: CRISPR, Expressing, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Comparative antioxidant activity of pomegranate leaf extract and derived metabolites in single and co-cultured cell lines

doi: 10.1038/s41598-025-20130-6

Figure Lengend Snippet: Schematic representation of the antioxidant activity of pomegranate leaf extract and derived metabolites in single and co-cultured cell lines. Primary antioxidant activity was evaluated by simultaneously incubating the pomegranate leaf extract (TE) and derived metabolites (EA and LU) with the pro-oxidant agent H 2 O 2 for 5 h. LU in HDF and EA in HUVEC resulted in a significant decrease in ROS-mediated cytotoxicity, thereby acting as primary antioxidants. Secondary antioxidant activity was evaluated by exposing the pre-incubating cells with pomegranate leaf extract (TE) and derived metabolites (EA and LU) for 24 h, followed by exposure to the pro-oxidant agent H 2 O 2 for 5 h. TE in the HDF-HUVEC resulted in a significant decrease in the ROS-mediated cytotoxicity, along with a significant increase in the mRNA expression of genes involved in the Nrf2 pathway, thereby acting as a secondary antioxidant.

Article Snippet: The human umbilical endothelial, HUVEC, cell line (C122-03, Merck), was cultured in the Endothelial Cell Growth Medium 2 (ECGM2, PromoCell, Italy) supplemented with the Supplement Mix (PromoCell) according to manufacturer’s instruction.

Techniques: Antioxidant Activity Assay, Derivative Assay, Cell Culture, Expressing