File Name : supplementary figure 1.docx

Caption : supplementary figure 1. experimental protocols in pine nut–sensitized mice. (a) ex vivo splenocyte experiments. six-week-old female c57bl/6 mice (n = 5) were orally sensitized with pine nut protein (pi; 5 mg protein/mouse) plus cholera toxin (ct; 10 μg) for 3 consecutive days during the first week, and once per week during the following 5 weeks. mice were euthanized and spleens were collected for splenocyte culture and stimulation. (b) in vivo oral treatment and challenge. mice (n = 5/group) were sensitized as in (a), while sham-sensitized controls received ct only. one week after the last sensitization dose, pi-sensitized mice were orally treated three times per week for 3 weeks with pbs, pi (5 mg protein/mouse), or the trypsin hydrolysate (t; 5 mg protein-equivalent/mouse), as indicated. three days after the last treatment dose, all groups were orally challenged with pi (50 mg protein/mouse). anaphylactic responses were assessed by rectal temperature and clinical score 30 min after challenge; blood was collected for pi-specific ige/igg1 and mcp-1 analyses.

File Name : supplementary figure 2.docx

Caption : supplementary figure 2. sds-page patterns of pine nut protein isolate (pi) and its hydrolysates obtained using pepsin (p), trypsin (t), alcalase (a), pepsin/trypsin (pt), pepsin/alcalase (pa), trypsin/alcalase (ta), and pepsin/trypsin/alcalase (pta). m: molecular weight marker. full, uncropped blots including all lanes and molecular mass markers are provided in supplementary material. all samples and controls used for comparative analysis were run on the same gel. cropping was applied only for clarity, and lane boundaries are indicated.