pore polycarbonate transwell inserts  (Corning Life Sciences)

Journal: Nature Communications

Article Title: Unlocking tumor barrier: annexin A2-mediated transcytosis boosts drug delivery in pancreatic and breast tumors

doi: 10.1038/s41467-025-61434-5

Figure Lengend Snippet: a The impact of protein pre-coating on nanocarrier transcytosis activity was studied using a transwell assay, wherein enhanced transcytosis activity of A2, A3, A5 and VTN pre-coating compared with albumin pre-coating particles and non-coated LC-MSNP ( n = 3 independent experiments, two-tailed unpaired t test) ( Created in BioRender. Meng, H. (2025) https://BioRender.com/bnpp34l ). b IVIS imaging revealed the biodistribution within the tumor and major organs ( n = 4 mice, two-tailed unpaired t test). Semi-quantification of fluorescent intensity unveiled a ~2-fold enrichment in KPC tumor distribution in the A2 coating group, without disrupting biodistribution in other organs. c The A2 coating approach was replicated in additional orthotopic models, namely breast cancer (EMT6, Py8119, and 4T1) and colon cancer (CT26) ( n = 4 mice, two-tailed unpaired t test). Encouragingly, ~6-fold improvement in tumor access was observed in the EMT6 BC model after a single IV injection. d A gold-core labeled version of F1, pre-coated with A2 protein, was prepared ( n = 3 independent experiments). e Tumor samples were harvested at 1- and 6-hour post IV injection, followed by tumor tissue TEM study. EC endothelial cells, VVO vesiculo-vacuolar organelle. Inserted images confirmed the presence of gold labeling within the particles. Green arrows: Transcytosing nanoparticles inside EC vesicles ( n = 3 independent experiments). f TEM image substantiated nanoparticle presence within a cancer cell 6 h post IV injection. Data are depicted as mean ± SD. Statistical significance was evaluated via two-tailed unpaired t test. Source data are provided as a Source Data file.

Article Snippet: HUVEC cells were seeded on the upper surface of the membrane in rat tail collagen-coated polycarbonate Transwell inserts (No. 3401 Costar; Corning; 0.4 μm pore size; 12 mm diameter) with a density of 5 × 10 5 cells per well.

Techniques: Activity Assay, Transwell Assay, Two Tailed Test, Imaging, IV Injection, Labeling

Journal: Nature Communications

Article Title: Unlocking tumor barrier: annexin A2-mediated transcytosis boosts drug delivery in pancreatic and breast tumors

doi: 10.1038/s41467-025-61434-5

Figure Lengend Snippet: a The CG model’s initial state (0 μs) showed random protein position and orientation relative to the lipid. After 50 μs of MD simulation with Martini force field, the equilibrated state was attained, characterized by protein adherence to the lipid membrane nano surface. b The Cα root mean square deviation (RMSD) of the protein concerning its initial positions within the CG model is depicted after 50 μs of simulation (upper panel) and further across three 500 ns simulations in the All-Atom (AA) model (lower panel). RMSDs were computed with backbone of protein alignment. c Time-Resolved contact area analysis illustrated the dynamic evolution of contact area between A2 and the lipid membrane over the entirety of the simulation. d Lipid surface-A2-α5β1 integrin interaction. Proteins are depicted in cartoon representation, with the lipid surface rendered in opaque gray. Notably, the α11 helix (red color) of A2 binds to the lipid, while helices (purple color) of A2 interact with α5β1 integrin. e While the interaction between A2 and α5β1 integrin was governed by hydrophobic interaction ( e 1), the interaction between A2 and nano surface (lipids) is mediated by electrostatic interaction ( e 3). The specific amino acids and their location in the indicated protein were demonstrated in the inserted boxes, e 2 (for A2 and α5β1 integrin) and e 4 (for A2 and lipids). f The HUVEC cell with integrin α5 subunit knockdown was detected by transwell for nanoparticle transcytosis assay ( n = 4 independent experiments, two-tailed unpaired t test)). g We mutated six amino acid residues on A2, replacing them with alanine. The resulting A2 mutant (A2-M) was used to pre-coated particles with NIR labeling, followed by the nanoparticle distribution study in an orthotopic EMT6 mouse model. A2 coating was used as control ( n = 5 mice, two-tailed unpaired t test)). h In the “zombie” mice, the blood vessels of EMT6 tumor-bearing mice were fixed, and a pump was employed to circulate blood containing A2-coated LC-MSNP. Data are depicted as mea n ± SD. Statistical significance was evaluated via two-tailed unpaired t-test ( n = 4 mice). Source data are provided as a Source Data file.

Article Snippet: HUVEC cells were seeded on the upper surface of the membrane in rat tail collagen-coated polycarbonate Transwell inserts (No. 3401 Costar; Corning; 0.4 μm pore size; 12 mm diameter) with a density of 5 × 10 5 cells per well.

Techniques: Membrane, Knockdown, Two Tailed Test, Mutagenesis, Labeling, Control

Journal: Nature Communications

Article Title: Host albumin redirects Candida albicans metabolism to engage an alternative pathogenicity pathway

doi: 10.1038/s41467-025-61701-5

Figure Lengend Snippet: A Growth of different C. albicans isolates and deletion mutants in RPMI-1640 with or without albumin assessed by optical density measurements every 30 min for 45 h at 600 nm ( n = 3). B Representative microscopic images showing SC5314, efg1 ΔΔ/ cph1 ΔΔ, and 101 grown with and without albumin in the absence of host cells. Images were taken after 24 h at ×10 magnification (scale bar = 200 µm). C Cytotoxicity of A-431 cells infected with C. albicans strains at different multiplicities of infections (MOI) after 45 h post infection (hpi) with or without albumin ( n = 3). D Comparison of cytotoxicity induced to A-431 cells in a transwell system (indirect infection) to directly infected A-431 cells in presence or absence of albumin after 45 hpi ( n = 4). E Cytotoxicity of A-431 cells infected with albumin pre-incubated or non-albumin pre-incubated C. albicans strains. The pre-incubation took place for 0.5 h, 1 h, 2 h, or 3 h. The cytotoxicity measurements were carried out after 45 hpi, as a control served a direct infection with or without albumin ( n = 4). Cytotoxicity ( C – E ) was quantified by measuring the LDH activity in the supernatant. Bars represent the mean ± SEM, and dots represent the mean of the technical replicates of the individual experiments. Data were tested for significance using a two-way ANOVA with Holm-Šídák multiple comparisons test ( C – E ). P values are provided in the figure for significant comparisons. Source data are provided as a Source Data file.

Article Snippet: Transwell inserts (polycarbonate membrane inserts with 0.4 μm pore size; Corning) were placed in the well and filled with 250 μL of the desired C. albicans strain (1 × 10 5 yeast cells/transwell) in RPMI-1640 medium with or without 10 mg/mL albumin.

Techniques: Infection, Comparison, Incubation, Control, Activity Assay