mle 12 mouse alveolar epithelial cells  (ATCC)

Journal: Biochemistry and Biophysics Reports

Article Title: Dual knockdown of Alox15 and TGF-β1 by lipid nanoparticle-delivered siRNA in bleomycin-induced pulmonary fibrosis

doi: 10.1016/j.bbrep.2026.102534

Figure Lengend Snippet: LNP-mediated delivery of siRNAs into lung epithelial and fibroblast cells effectively reduces mRNA levels both in vitro and in vivo . (A, B) MLE-12 cells and murine lung fibroblasts were treated with negative control (NC), Alox15, or TGF-β1 siRNA for 24 h, followed by treatment with bleomycin for an additional 24 h, then subjected to real-time PCR assay. Fluorescence images of (C) MLE-12 cells and (D) murine lung fibroblasts treated with 50 nM naked siRNA-Cy5 or siRNA-Cy5 encapsulated with LNPs (siRNA@LNP) for 6 h, followed by staining with Hoechst 33342. Scale bar = 20 μm. (E) Flow cytometry analysis showing the mean fluorescent intensity (MFI) of Cy5 positive cells. (F) Mice were injected with saline, naked siRNA, or siRNA@LNP and sacrificed at 24 h after treatment. Cy5 fluorescence signal in the heart, lung, liver, spleen and kidney was measured by The PhotonIMAGER Optima system immediately. Fluorescence intensity in the lung expressed as total radiant efficiency. n = 3 per group. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Scatter plots representing individual mice.

Article Snippet: Murine lung epithelial cells (MLE-12 cells) were obtained from ATCC (Manassas, VA, USA) and cultured in DMEM/F12 medium (Corning) containing 4% FBS, 0.005 mg/mL insulin, 0.01 mg/mL transferrin, 30 nM sodium selenite, 10 nM hydrocortisone, 10 nM β-estradiol, 10 mM HEPES, 2 mM l -glutamine, 100 U/mL penicillin G, and 100 μg/mL streptomycin.

Techniques: In Vitro, In Vivo, Negative Control, Real-time Polymerase Chain Reaction, Fluorescence, Staining, Flow Cytometry, Injection, Saline