Journal: bioRxiv
Article Title: Kinetics of local C3 production orchestrates neutrophil recruitment in lung injury
doi: 10.64898/2026.02.17.706326
Figure Lengend Snippet: ( A ) Schematic of infection in mouse model. Created in BioRender. Kulkarni, H. (2026) https://BioRender.com/jevlom7 ( B-C ) Assay for detecting C3 activation on lipopolysaccharide (LPS) normalized to wildtype (WT) uninfected ( B ) bronchoalveolar lavage (BAL) and ( C ) serum of WT mice infected with Pseudomonas (Pa) and euthanized at 2, 4, 8, 12 and 24 h. Results for ( D ) full-length C3 ELISA and ( E ) C3a-neoepitope ELISA in BAL post- Pa infection. ( F ) Schematic of infection in ex vivo human lung model. Modified based on Zhang et al. ( G-I ) Changes in BAL ( G ) C3, ( H ) C3b/iC3b, and ( I ) IgM levels in ex vivo human lung pre-infection (T0, 0 h) and post-infection (T4, 4h). Distribution of data is represented as scatterplots showing individual data points, and bars represent means ± SD. ****P<0.0001, ***P < 0.001, **P < 0.01, and *P < 0.05 using ordinary one-way ANOVA after adjusting for multiple group comparisons, and Mann-Whitney test for two-group comparison. Abbreviations: CFU: colony-forming units; FACS: fluorescence-associated cell sorting (flow cytometry); PEEP: positive end-expiratory pressure; RR: respiratory rate; VT: tidal volume.
Article Snippet: For use with BAL and serum, 96-well flat-bottom ELISA plates (#3855, Thermo Fisher Scientific) were coated with a C3b capture antibody (1:50 dilution; 100 μl per well in PBS; #HM1045, rat anti-mouse C3b/iC3b/C3d/C3dg, clone 11H9; Hycult Biotech) and incubated overnight at 4°C.
Techniques: Infection, Activation Assay, Enzyme-linked Immunosorbent Assay, Ex Vivo, Modification, MANN-WHITNEY, Comparison, Fluorescence, FACS, Flow Cytometry