Journal: NPJ Precision Oncology

Article Title: Pulmonary surfactant protein D reduces lung cancer progression associated with decreased IL-4/STAT6 signaling

doi: 10.1038/s41698-025-01163-6

Figure Lengend Snippet: a A549 cells were treated with rhSP-D (10 mg/ml) for 2 h, followed by incubation with recombinant human IL-4 (10 ng/ml) or IL-13 (10 ng/ml) for 15 min before harvest. Protein expression and phosphorylation were assessed by western blotting. b Interleukin 4 receptor α (IL-4R-α) and BSA were coated (1 mg/ml) in 96 ELISA plates and different concentrations of rhSP-D were added. After 2 h, the relative binding was evaluated by colourimetic ELISA assay (optical density (OD)). c Representative surface plasmon resonance fits of binding of rhSP-D in the concentration range 1.5–200 nM to immobilized IL-4R-α. Experimental data, expressed as resonance units (RU), are shown as solid lines and fitted curves as dotted lines. The following parameters were determined; k a = 7.1 ± 1.9 × 10 3 /Ms, k d = 1.7 ± 0.4 × 10 −4 /s, K D = 25 ± 5 nM. Average chi 2 / R max of the fits was 0.011. d Dual injection on the IL-4R sensor chip. First injection with 100 nM rhSP-D followed by addition of either 5 mM CaCl 2 (black), 10 mM EDTA (purple), 100 mM mannose (blue), or 100 mM N-acetyl-D-glucosamine (GluNac) (red). e Association-phase of the injection of 100 nM rhSP-D followed by addition of either 5 mM CaCl 2 (black), 10 mM EDTA (green), 1–100 mM mannose (blue) or 1–100 mM N-acetyl-D-glucosamine (GluNac) (red).

Article Snippet: A549 cells were treated with rhSP-D (50 μg/ml) for 2 h. Following, the cells were stimulated by recombinant human IL-4 (R&D systems, 6507-IL) or recombinant human IL-13 (R&D systems, 213-ILB) for 15 min.

Techniques: Incubation, Recombinant, Expressing, Phospho-proteomics, Western Blot, Enzyme-linked Immunosorbent Assay, Binding Assay, SPR Assay, Concentration Assay, Injection