P dimethylaminobenzaldehyde

Silpot 184 silicone elastomer was purchased from Dow Corning Toray Co., Ltd. Acetone, ethanol (95), ethanol (99.5), polyethylene glycol 2000 (PEG2000), heptane, hydrochloric acid, sodium hydroxide, methyl orange, p-dimethylaminobenzaldehyde, and hydrazine dihydrochloride were obtained from FUJIFILM Wako Pure Chemical Industries (Osaka, Japan). An ABS filament was obtained from WANHAO.

The inhibitory activity on hyaluronidase was measured according to a modified Morgan–Elson method as described previously [16 (link),17 (link)]. Briefly, samples prepared in 0.1 M acetate buffer (pH 4) and hyaluronidase (type IV-S: From bovine testes, Sigma, St. Louis, MO, USA) in buffer with a final concentration of 5 mg/mL were incubated at 37 °C. Then, compound 48/80 (Sigma) in buffer with a final concentration of 0.5 mg/mL was added and incubated. After hyaluronic acid sodium salt (from rooster comb, Wako, Osaka, Japan) in buffer with a final concentration of 0.4 mg/mL had been included, the mixture was incubated. p-Dimethylaminobenzaldehyde (Wako) acetate solution was then added and incubated. Then the absorbance was determined at 585 nm, and the enzyme inhibitory activity (%) was quantified. EsB or Esg-B as a stock solution (2.8 mg/mL) was prepared with 0.7% DMSO and diluted with 0.1 M acetate buffer. Disodium chromoglycate (Wako) and suplatast tosylate (TCI, Tokyo, Japan) were prepared with water as a stock solution of 10 mg/mL.

The chemicals used were as follows: bleomycin hydrochloride, heparin, L-hydroxyproline, paraformaldehyde, Triton X-100, formamide, p-dimethylaminobenzaldehyde, 1-propanol, and perchloric acid (Wako Pure Chemical, Japan); Evans blue and chloramine T (Sigma, MO); OCT compound (Sakura Finetek, Japan); DAPI (Dojindo Laboratories, Japan), random 9 mers and Ex Taq DNA polymerase (Takara Bio, Japan); ReverTra Ace (Toyobo, Japan); ethidium bromide, goat anti-rabbit IgG (H+L) Alexa Fluor 488 (#A-11008, Invitrogen, CA), and goat anti-mouse IgG (H+L) Alexa Fluor 568 (#A-11008, Invitrogen, CA); rabbit anti-H-PGDS polyclonal antibody (#O60760, Cayman Chemical, MI); mouse anti-CD68 monoclonal antibody (#MCA341R, AbD Serotec, UK); mouse anti-Gr-1 antibody (#108413, Biolegend, CA).