Anti type 1

In the mouse experiment, sections were incubated with the primary antibody anti-type II collagen (Daiichi Fine Chemical). In the rabbit experiment, sections were incubated with the primary antibodies anti-type I (abcam) and anti-type II collagen (Fuji Pharm.). The samples were incubated with a biotinylated secondary antibody and visualized by the precipitation of diaminobenzidine (DAKO). For CCL21 and CCR7 evaluation, sections were incubated with the primary antibodies anti-CCL21 (R&D), anti-CCR7 (Abcam), and staining was visualized with Alexa Fluor 568 (Invitrogen), FITC Conjugated AffiniPure (Jackson Immuno Research). Nuclei were stained with DAPI (Invitrogen).

Cells were fixed for 10 min at − 30 °C with cold methanol, and blocked with 1% FBS in PBS for 60 min at room temperature. The cells then were stained with the antibodies anti α-SMA (1/2000) (Sigma-Aldrich), anti-Type I (1/100), and IV (1/100) collagen (Abcam). As secondary antibodies, Alexa Fluor 594 Goat Anti-Mouse IgG (1/1000) and Alexa Fluor 488 Goat Anti-rabbit IgG (1/1000) (Thermo Fisher Scientific) were used. Cell nuclei were then stained with DAPI (5 μg/mL) (Dojindo, Kumamoto, Japan) and the cells were inspected with the fluorescence microscope BZ9000 (Keyence, Osaka, Japan).