Fig. 5. Autoantigenic substrate cleavage by MMP-9.

a Left: incubation of PNMA2 (1 µM) alone or with active MMP-9 at different enzyme/substrate (E/S) ratio’s (1/10, 1/100, 1/1000, and 1/10 000) overnight at 37 °C, shows cleavage into PNMA2 fragments after SDS-PAGE separation and silver staining. Right: incubation of PNMA2 with active MMP-9 (E/S ratio of 1/10) for 0, 5, 10, 30, 90, 180, 420 min and overnight. b Similar experimental set up for VIP (10 µM), c Similar experimental set up for Ri (0.2 µM). d Recoverin (10 µM) was not cleaved after prolonged incubation at an enzyme/substrate ration of 1/10. For all panels molecular weight standardization was included and the molecular weight marker proteins are indicated in kilodaltons (kDa). All photographs are aligned in such ways that for all cleavage experiments the conditions were similar to the ones indicated in the top photographs of panel A