Journal: bioRxiv

Article Title: Progressive matrix stiffening of tyramine-modified silk fibroin hydrogels governs stage-specific pulmonary fibroblast activation

doi: 10.64898/2026.06.01.729382

Figure Lengend Snippet: A) Hydrogels were composed of either 0% or 50% mixture of tyramine-modified silk and were exposed to media without or with 5 ng/mL TGFβ. After hydrogels were cast, 25,000 NHLF cells were seeded on the surface. B) SEM imaging of the surface of the silk hydrogels incubated without cells for Day 0 and Day 14. C) Live/Dead imaging of Calcein AM for live cells (green) and Ethidium Homodimer-1 for dead cells (red) indicated cytocompatibility. D) Alamar Blue Assay for metabolic activity indicated that 0% gels showed an increase in activity over 14 days, while 50% gels plateaued from day 7 to 14. E) Mammalian β-Galactosidase Assay for senescence revealed no change for any timepoints tested over 14 days. N=3-4, Error bars = SEM.TCP = Tissue Culture Plate, T-= without TGFβ, T+ = 5 ng/mL of TGFβ. Two-way ANOVA, Tukey’s: ns = not significant p > 0.05, * = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001.

Article Snippet: Normal human lung fibroblasts (NHLFs, PCS-201-013, ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle Medium, high glucose (DMEM, ThermoFisher) supplemented with 10% Gibco Fetal Bovine Serum, Premium (FBS, ThermoFisher), 1% Gibco non-essential amino acids (ThermoFisher), and 1% Gibco antibiotic–antimycotic (ThermoFisher).

Techniques: Modification, Imaging, Incubation, Alamar Blue Assay, Activity Assay

Journal: bioRxiv

Article Title: Progressive matrix stiffening of tyramine-modified silk fibroin hydrogels governs stage-specific pulmonary fibroblast activation

doi: 10.64898/2026.06.01.729382

Figure Lengend Snippet: Top row: NHLFs on 0% SF-TA hydrogels at Day 2, Day 7, and Day 14, exhibiting a proliferative, inflammatory phenotype consistent with early fibroblast activation, characterized by secretion of IL-6, IL-8, and MCP-1 (dots) and progressive collagen deposition (fibers). Bottom row: NHLFs on 50% SF-TA hydrogels at Day 2, Day 7, and Day 14, exhibiting metabolic plateau and lower inflammatory cytokine secretion, gradual collagen secretion, with TGFβ-dependent myofibroblast activation. Together, these two substrate conditions recapitulate distinct stages of the fibrotic disease continuum.

Article Snippet: Normal human lung fibroblasts (NHLFs, PCS-201-013, ATCC, Manassas, VA, USA) were cultured in Dulbecco’s Modified Eagle Medium, high glucose (DMEM, ThermoFisher) supplemented with 10% Gibco Fetal Bovine Serum, Premium (FBS, ThermoFisher), 1% Gibco non-essential amino acids (ThermoFisher), and 1% Gibco antibiotic–antimycotic (ThermoFisher).

Techniques: Activation Assay

Journal: Cell Death Discovery

Article Title: Trifluoperazine causes mast cell apoptosis through a secretory granule-mediated pathway

doi: 10.1038/s41420-026-03122-x

Figure Lengend Snippet: A Human lung smooth muscle cells (HLSMCs), lung fibroblasts (HLFs), and small airway epithelial cells (HSAECs) were treated with TFP at the indicated concentrations for 24 h. Human peripheral blood eosinophils and neutrophils were treated with TFP for 2 h. Cell viability was assessed by staining the cells with Annexin V (AnnV) and DRAQ7. Viable cells, AnnV − DRAQ7 − ; apoptotic cells, AnnV + DRAQ7 − ; necrotic/late apoptotic cells, AnnV + DRAQ7 + . HSAECs, n = 6 from three independent experiments; HLSMCs, eosinophils, neutrophils, n = 4 from four independent experiments; HLFs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA for HLSMCs, HLFs; Kruskal-Wallis for HSAECs; Friedman test for eosinophils, neutrophils). B – C Bone marrow-derived MCs (BMMCs) and peritoneal cell-derived MCs (PCMCs) treated under the same conditions as in ( A ) for 24 h. BMMCs, n = 5 from two independent experiments; PCMCs, n = 3 from one individual experiment representative of three independent experiments (One-way ANOVA). Untreated (control) cells were used for statistical comparisons to all other groups in all figures. The bar charts show mean values + SEM or median + interquartile range. * P < 0.05; ** P < 0 .01; **** P < 0.0001. D Effect of TFP on DNA degradation. MCs were preincubated with bafilomycin A1 (Baf A1) (20 nM) for 2 h followed by treatment with TFP (10 μΜ) for 2 h. DNA was extracted from MCs and fragmentation was assessed by agarose gel electrophoresis. St standard marker.

Article Snippet: Primary human lung fibroblasts (HLFs) (PCS-201-013) and primary human lung smooth muscle cells (HLSMCs)(PCS-130-010) were obtained from American Type Culture Collection (ATCC, Manassas, VA) and cultured following the manufacturer’s instructions.

Techniques: Staining, Derivative Assay, Control, Agarose Gel Electrophoresis, Marker