Figure 3

Isolation and detection of active forms of TGF-β, MMP2 and MMP11 in porcine dental pulp. (a) Heparin-Sepharose chromatogram of extracts from porcine dental pulp detected at 280 nm absorbance. Downward-pointing arrows represent the starting points of the step gradient with 0.05, 0.1 and 0.2 M NaCl. (b) ALP-inducing activity of HPDL cells with (+) or without (−) the addition of SB431542 to fractions P1-P4 isolated by chromatography. Recombinant human TGF-β1 with a carrier (0.3 ng mL⁻¹) (TGF-β1) was used as a positive control for the detection of ALP-inducing activity in HPDL cells. The data show the average value of six measurements. (c) A gelatin zymogram of fractions P1-P4 incubated without (left) or with (right) EDTA. (d and e) Western blots of fractions P1-P4 with specific antibodies against (d) MMP2 and (e) MMP11. The full-length gel (c) and blots (d and e) are presented in Supplementary Fig. S4.