Journal: Nature biomedical engineering

Article Title: Collagen-binding IL-12 enhances tumour inflammation and drives the complete remission of established ‘immunologically cold’ murine tumours

doi: 10.1038/s41551-020-0549-2

Figure Lengend Snippet: a, Schematic illustrating the fusion sites of the von Willebrand factor A3 CBD to murine p35 and p40 subunits. CBD was fused to each of the subunits via a (GGGS)2 linker. b, Dose-response relationship of phosphorylated STAT4 with IL-12 and CBD-IL-12 in preactivated primary mouse CD8+ T cells (n = 3, mean ± SD). c,d, Affinity (KD values are shown) of CBD-IL-12 against collagen I (c) and collagen III (d) as measured by SPR. CBD-IL-12 was flowed over the chips at indicated concentrations. Curves represent the specific responses (in resonance units, RU) to CBD-IL-12. Experimental curves were fitted with 1:1 Langmuir model. Dissociation constants (KD) and rate constants (kon and koff) determined from the fitted curves are shown. e,f, Binding of unmodified IL-12 (e) or CBD-IL-12 (f) to human melanoma cryosections was imaged by fluorescence microscopy. Scale bars, 100 μm. Experiments were performed twice (b,c,d,e,f), with similar results. Representative data are shown.

Article Snippet: Tissues were then stained with the following primary antibodies: rabbit anti-human collagen I antibody (Abcam, ab34710), mouse anti-human CD31 antibody (Abcam, ab119339) and rat anti-mouse IL-12p70 antibody (from mouse IL-12p70 ELISA kit, Invitrogen).

Techniques: Binding Assay, Fluorescence, Microscopy