Fig. 4.
Gingipain profile of P. gingivalis sialidase mutants compared with vimA mutant (FLL92) and wild-type W83 from exponential-phase cultures. Acetone-precipitated proteins isolated from supernatants from cultures grown to an OD600 of 0.8 were analyzed by SDS-PAGE and stained with SimplyBlue SafeStain (A) or analyzed by immunoblotting using gingipain-specific antibodies (B to D). Approximately 20 to 25 μg of protein was loaded in each lane. Antibodies: chicken anti-RgpA (B), rabbit anti-RgpB (C), and chicken anti-Kgp (D). Ex, extracellular fraction; CL, cell lysate fraction. Arrows indicate the unprocessed, partially processed, or mature gingipains; MW, molecular weight markers (thousands).