Anti col 1

For immunohistochemical staining, paraffinized sections (5 μm) of PDLSC or JBMSC sheets were deparaffinized; blocked; and incubated for 1 h in the following primary antibodies: (1) anti-FN1 (1:200), (2) anti-COL I (1:200), and (3) anti-TNALP (1:200) (1:200, all from Santa Cruz Biotechnologies, Dallas, TX, USA). PBS was used in place of the primary antibodies for negative controls. The sections were then incubated for 45 min in biotinylated secondary antibodies (1:1000) purchased from ZSGB BIO (Peking, China). Each experiment was repeated in triplicate.

The harvested cell aggregate, native and decellularized tissue sections from porcine heart and TA muscle from rat were fixed in 4% phosphate-buffered paraformaldehyde for 24 hours, embedded in paraffin. Eight-micrometer-thick serial sections were cut from the paraffin-embedded blocks and underwent H&E staining and Masson’s Trichrome staining. Immunohistochemical staining was performed using standard procedures as described previously [35 (link), 36 (link)]. Briefly, sections were incubated with primary antibodies as follows: anti-Col-I (1:200; Santa Cruz, USA), anti-fibronectin (1:200; Abcam) and anti-integrin-β1 (1:200; Abcam). The same source IgG was used for the negative control instead of the primary antibodies. Biotinylated secondary antibodies (1:1000) were purchased from Sigma-Aldrich. The stained sections were observed using the light microscope (Nikon, Japan). The photographs were evaluated by Image-Pro Plus 6.0 (Media Cybernetics, USA) from three randomly selected views of each specimen.