murine lung 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

Journal: Signal Transduction and Targeted Therapy

Article Title: A nanobody-based proteolysis-targeting chimera offers broad-spectrum protection against diverse influenza virus infections

doi: 10.1038/s41392-026-02666-9

Figure Lengend Snippet: Antiviral efficacy of Nb-PROTACs against H1N1 and H5N1 viruses in mice. a Antiviral activity of mVHL-Nb170 in MLE-12 cells. Cells stably expressing Nb170 or mVHL-Nb170 were infected with SD012 at an MOI of 0.01. The supernatants were collected at the indicated time points and subjected to viral titration in MDCK cells. The data are presented as the means ± SDs from three independent biological samples ( n = 3). b Schematic representation of the experimental design for the in vivo study. BALB/c mice received two intratracheal doses of AAV-LungM3 at an interval of 3 days, each at 5 × 10 10 vg, resulting in a total dose of 1 × 10 11 vg per mouse. Three weeks post-administration, the mice were intranasally challenged with 5 MLD 50 of the PR8 (H1N1) or SD012 (H5N1) virus. c , d Protective efficacy of mVHL-Nb170 in mice. Groups of 14 mice were administered the AAV-LungM3 vector expressing the indicated proteins and subsequently challenged intranasally with 5 MLD 50 of PR8 ( c ) or SD012 ( d ). Nasal turbinates and lungs were collected on day 3 postchallenge for viral titration. The data are presented as the means ± SDs from four animals ( n = 4). Survival and body weight were monitored daily for 14 days ( n = 10). The percentages in c and d denote the mean decrease in viral load compared with that in the controls. Asterisks indicate significant differences (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The human embryonic kidney cell line 293T, murine lung epithelial cell line MLE-12, and human lung epithelial cell line A549 were obtained from the American Type Culture Collection (ATCC).

Techniques: Activity Assay, Stable Transfection, Expressing, Infection, Titration, In Vivo, Virus, Plasmid Preparation

Journal: Cancer Research

Article Title: ACSL5 Mediates Adaptation to the Palmitic Acid–Enriched Pulmonary Microenvironment to Enhance Metastatic Breast Cancer Cell Survival and Lung Metastasis

doi: 10.1158/0008-5472.CAN-25-0866

Figure Lengend Snippet: Exo-derived USP47 mediates fatty acid synthesis in AT2. A, MLE-12 cells were cocultured with exosomes derived from the control (shNC) and USP47-knockdown (shUSP47) MDA-MB-231/LM3. YAP, FASN and ACLY proteins, as well as the PA levels ( n = 5), in MLE-12 cells were detected. B, MLE-12 cells were cocultured with exosomes derived from the control (vector) and USP47-overexpression (oeUSP47) MDA-MB-231/PRI. YAP, FASN, and ACLY protein levels, as well as the PA levels ( n = 5), in MLE-12 cells were detected. C, Left, Western blotting was performed to detect YAP, FASN, and ACLY protein levels in the control (shNC) or YAP-knockdown (shYAP) MLE-12. Right, relative PA levels ( n = 5) in MLE-12 cells were detected. D, MLE-12 cells were cocultured with the control exosomes (vector) or USP47-overexpression exosomes (oeUSP47) derived from MDA-MB-231/PRI, or YAP-knockdown MLE-12 cells and its controls were cocultured with USP47-overexpression exosomes (oeUSP47) derived from MDA-MB-231/PRI. FASN and ACLY protein levels, as well as the PA levels ( n = 5), in MLE-12 cells were detected. E, MLE-12 cells were cocultured with the control exosomes (shNC) or USP47-knockdown exosomes (shUSP47) derived from MDA-MB-231/LM3, or YAP-overexpression (oeYAP) and its control MLE-12 cells (Vector) were cocultured with USP47-knockdown exosomes (shUSP47) derived from MDA-MB-231/LM3, respectively. FASN and ACLY protein levels, as well as the PA levels ( n = 5), in MLE-12 cells were detected. F and G, Relative PA levels in MLE-12 in indicated group ( n = 5). H, Relative PA levels ( n = 5) in the lung interstitial fluid of BALB/c nude mice injected with the control (shNC) or USP47-knockdown (shUSP47) MDA-MB-231/LM3 cells. I, Representative BLI and metastatic lungs of BALB/c nude mice injected with the control (shNC) or USP47-knockdown (shUSP47) MDA-MB-231/LM3 cells. J, Quantification of BLI and lung metastatic nodules ( n = 7). All data represent the mean ± SD. One-way ANOVA ( A , C–G , H , and J ) and Student t test ( B ) were utilized. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: Triple-negative breast cancer cell lines 4T1, MDA-MB-231, and HCC1806 and mouse alveolar epithelial cell line MLE-12 were purchased from the ATCC.

Techniques: Derivative Assay, Control, Knockdown, Plasmid Preparation, Over Expression, Western Blot, Injection