mle 12 cell line  (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: bioRxiv

Article Title: Cryptochrome Loss Drives COPD-like Lung Pathology through Disrupted Alveolar Epithelial Proliferation and Immune Homeostasis

doi: 10.64898/2026.05.19.726266

Figure Lengend Snippet: MLE12 cells were transfected with 50 pmole/ml control, Cry1 , or Cry2 siRNA on day 0 and day 1. A. Western blot showed CRY1/CRY2 protein levels on day 2. B. The transcript levels of cytokines, including C-C motif chemokine ligand 2 ( Ccl2 ), C-X-C motif chemokine ligand 1 ( Cxcl1 ), Interleukin-6 ( Il6 ), Il8/Cxcl5 , matrix metalloproteinase 1 and 7 ( Mmp1 and Mmp7 ), and Tnfa were determined with RT-qPCR. C. Cell numbers were measured with the WST1 assay 1-, 2-, or 3-days post siRNA transfection. n=6. D-E. Primary alveolar ECs were isolated from WT or Cry1/2 dKO mice for RNA sequencing analysis. D. Heatmap image of up (501) and down (369) regulated genes in Cry1/2 dKO mice alveolar ECs compared to WT alveolar ECs. Each gene is represented as a horizontal line. E. GO Biological Process (BP) analysis of DEGs that were in Cry1/2 dKO lung ECs. F. GO Molecular Function (MF) analysis of DEGs. G. GO Cellular Component (CC) analysis of DEGs. The top 5 terms were shown for both up and down-regulated genes. H. Enriched pathways (Metascape) of overlapping genes from human COPD patient ECs DEGs with DEGs from (D). I. GO BP, J. MF and K . CC analysis of common genes between human COPD patient lungs with DEGs in Cry1/2 dKO ECs. Data are presented as mean ± SEM. ** p<0.1, *** p<0.001, and **** p<0.0001 Cry1/2 si vs si_con (Two-way ANOVA followed by Bonferroni adjusted multiple comparisons). For C , only the asterisks for si- Cry1/2 are shown.

Article Snippet: Human lung epithelial cell line A549 and mouse primary lung epithelial cell line MLE12 were procured from ATCC (Manassas, VA).

Techniques: Transfection, Control, Western Blot, Quantitative RT-PCR, Isolation, RNA Sequencing

Journal: bioRxiv

Article Title: Cryptochrome Loss Drives COPD-like Lung Pathology through Disrupted Alveolar Epithelial Proliferation and Immune Homeostasis

doi: 10.64898/2026.05.19.726266

Figure Lengend Snippet: A. Experimental schematic: Cry1/2 dKO mice were fed with regular (RD) or 1% NOB-containing diets starting at 1 month old and analyzed at 4 months. B. Representative images of H&E-stained lung sections collected from mice fed with a RD or NOB diet. C. MLI quantification of H&E-stained lung sections from male or female Cry1/2 dKO fed with RD or NOB diets. D. RNA-seq was performed using primary lung ECs isolated from the WT mice, or Cry1/2 dKO mice fed with RD or NOB diet. Heatmap of RNA-seq data of lung ECs from different treatment groups. E. GO BP analysis of DEGs that were rescued by NOB treatment in Cry1/2 dKO lung ECs. F. GO MF analysis of rescued DEGs. G. GO CC analysis of rescued DEGs. H. RT qPCR confirmed the upregulation and rescue of genes involved in cilium movement. I. The transcript expression changes of important differentially expressed chemokines, cytokines, Mmps , and surfactant proteins from RNA-seq data. Data were presented as fold changes to WT. J. MlE12 cells were transfected with control siRNA or siRNA targeting Cry1 and Cry2 . 36 h after transfection, cells were treated with 5 ng/ml TNF-α for 15 min. Phosphorylated P-65 and P65 levels were determined with Western blots. K. MLE12 cells were treated with 20 µM NOB for 24 h, followed by 5 ng/ml TNF-α for 0, 15, or 30 mins. Phosphorylated P-65 and P65 levels were determined with Western blots. L. MLE12 cells were transfected with control siRNA, or siRNA targeting Cry1 and Cry2 , and treated with 20 µM NOB for 24 h before TNF-α stimulation (15 mins). Representative confocal images of p65 (green) are shown (×400 magnification; scale bar = 10 µm). M. The percentage of cells with p65 localized in the cytoplasm (C), both cytoplasm and nucleus (C+N), or nucleus (N) was quantified for each treatment condition. N. Western blot of CRY1/2 and COPD-associated proteins in primary lung ECs from different treatment groups. Data are presented as mean ± SEM. P-value was calculated from a two-tailed t-test for H and I, * p<0.05, ** p<0.01, *** p<0.001, and **** p<0.0001 in RD vs in Cry1/2 dKO samples.

Article Snippet: Human lung epithelial cell line A549 and mouse primary lung epithelial cell line MLE12 were procured from ATCC (Manassas, VA).

Techniques: Staining, RNA Sequencing, Isolation, Quantitative RT-PCR, Expressing, Transfection, Control, Western Blot, Two Tailed Test

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