Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Neutrophilic granule protein promotes lipopolysaccharide-induced iNOS/NO expression via JAK2/STAT1 signaling pathway and augments bacteria clearance of macrophages

doi: 10.1007/s00018-025-05865-9

Figure Lengend Snippet: NGP controls the expression of iNOS/NO by regulating the JAK2/STAT1 signaling pathway. ( A ) Ngp NC /RAW and Ngp HI /RAW cells were treated with LPS (10 μg/mL) for 6 h and lysed for RNA sequencing. The KEGG database was utilized to enrich signal pathways exhibiting significant differences. ( B ) Ngp NC /RAW and Ngp HI /RAW cells were stimulated with LPS for 6 h. The cells were harvested and subjected to CSP100 PLUS phosphorylation microarray analysis. ( C ) The Ngp NC /RAW and Ngp HI /RAW cells were treated without or with LPS for 6 and 12 h respectively. Western blotting analysis was performed to assess the phosphorylation levels of JAK2 and STAT1. ( D ) The Ngp NC /RAW and Ngp HI /RAW cells were treated with LPS for 6 h and subjected for IP-MS to screen for the binding protein of NGP and verify the interaction. ( E ) The Ngp WT /RAW cells were treated without or with LPS for 6 h, harvested for Co-IP, and STAT1 expression levels were quantified via grayscale intensity analysis. ( F – H ) The predicted structures of NGP and STAT1 were generated by Alphafold and subjected to GRAMM Web Server for molecular docking. ( F ) Molecular docking generic matching information. ( G ) Predicted binding sites between NGP and STAT1. ( H ) The protein–protein interaction cartoon model generated by PyMOL. (I-J) The phosphorylation levels in PBS- or LPS-treated Ngp NC /RAW and Ngp HI /RAW cells were examined by LSCM (Scale bar = 20 μm). (K-M) Ngp NC /RAW and Ngp HI /RAW cells were pre-treated without or with JAK2 inhibitor (10 μM), STAT1 inhibitor (50 μM), or control solution for 2 h and then stimulated without or with LPS for 12 h. The supernatants and cells were harvested for NO expression, Nos2 mRNA level and positive ratio of iNOS by aforementioned methods ( n = 3). (N–O) Ngp NC /RAW and Ngp. HI /RAW cells were transfected with scramble siRNA or STAT1 siRNA, respectively, followed by 12-h stimulation with PBS or LPS. The supernatants and cells were harvested for concurrent measurement of NO levels and Nos2 mRNA expression via Griess assay and qRT-PCR ( n = 3). The data represent the mean ± SEM of at least three independent experiments. Statistical analysis was performed using one-way ANOVA to compare the results. * p < 0.05

Article Snippet: STAT1 antibody (bsm-33270 M) for LSCM was obtained from Bioss (Beijing, China).

Techniques: Expressing, RNA Sequencing, Phospho-proteomics, Microarray, Western Blot, Protein-Protein interactions, Binding Assay, Co-Immunoprecipitation Assay, Generated, Control, Transfection, Griess Assay, Quantitative RT-PCR

Journal: Materials Today Bio

Article Title: An integrin-based quercetin 7-rhamnoside liver-targeted delivery liposomes for intrahepatic cholestasis in pregnancy

doi: 10.1016/j.mtbio.2025.102031

Figure Lengend Snippet: Evaluation of hepatocyte and biliary epithelial cells targeting characteristics in vitro . (A) LSCM fluorescence images of Heap1-6 cell line after incubation with Free-C6, C6-Lip, and t-C6-Lip for 1, 2, and 4 h; (B) LSCM fluorescence images of HCCC-9810 cell line after incubation with Free-C6, C6-Lip, and t-C6-Lip for 1, 2, and 4 h; (C) Semi-quantitative fluorescence intensity analysis of the Heap1-6 and HCCC-9810 cell line uptake of Free-C6, C6-Lip, and t-C6-Lip for 1, 2, and 4 h ( n = 3); ∗, p < 0.05 ; ∗∗, p < 0.01 ; and ∗∗∗, p < 0.001 compared to the Free-C6 group.

Article Snippet: A Nikon LSCM was used to observe the cellular uptake and fluorescence distribution of DAPI and C6.

Techniques: In Vitro, Fluorescence, Incubation