Journal: Materials Today Bio
Article Title: Naringin nanoparticles alleviate RA-ILD pulmonary fibrosis by targeting 14-3-3ζ to inhibit LYVE1 + macrophages TGF-β1 secretion
doi: 10.1016/j.mtbio.2026.102982
Figure Lengend Snippet: NAR-induced LYVE1 + macrophages suppress EMT in pulmonary epithelial cells and FMT in lung fibroblasts. (A) CD68 + LYVE1 - BLM and CD68 + LYVE1 + BLM were sorted by flow cytometry. (B-C) After LYVE1 + BLM and LYVE1 - BLM were induced by 150 μM NAR for 48 h, analyzed the expression of LYVE1 and 14-3-3ζ mRNA and protein levels by RT-qPCR and western blotting (n = 3). (D) The flowchart of co-culture of NAR-induced BLM and A549/HFL1 cells. (E-F) Scratch experiment, added LYVE1 - BLM and LYVE1 + BLM which were induced by NAR, and set up Ctrl. Scale bar: 750 μm (n = 3). (G-H) Transwell invasion experiment, LYVE1 - BLM and LYVE1 + BLM incubated with 150 μM NAR for 48h were added to the lower chamber of 24-well plate and Ctrl was set, and A549 cells were added to the upper chamber. scale bar: 750 μm (n = 3). (I-J) LYVE1 - BLM/LYVE1 + BLM and A549 cells were co-cultured with 150 μM NAR for 48 h, and analyzed the expression of EMT-related (E-cadherin, vimentin) mRNA and protein levels of A549 cells by RT-qPCR and western blotting (n = 3). (K-L) LYVE1 - BLM/LYVE1 + BLM and HFL1 cells were co-cultured with 150 μM NAR induction for 48h, and analyzed the expression of FMT-related (fibronectin, COL1A1, α-SMA) mRNA and protein levels of HFL1 cells by RT-qPCR and western blotting (n = 3). (M) Representative images showing immunofluorescence staining for E-cadherin, vimentin, fibronectin, COL1A1 and α-SMA (RED) in mice lung tissue, scale bar: 100 μm, 50 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
Article Snippet: To conduct the staining procedure, employ the following antibodies: CD68 (santa cruz biotechnology, inc) and LYVE1 (fitzgerald industries international, LYVE1 antibody).
Techniques: Flow Cytometry, Expressing, Quantitative RT-PCR, Western Blot, Co-Culture Assay, Incubation, Cell Culture, Immunofluorescence, Staining