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. 2014 Mar 20;289(19):13531–13542. doi: 10.1074/jbc.M113.525287

FIGURE 2.

FIGURE 2.

Presence of PTX3 in both AM soluble extract and the purified HC-HA complex. A and B, recombinant human PTX3 control, purified human IαI control, AM extract (AME) and the AM HC-HA complex were treated with or without 50 mm NaOH at 25 °C for 1 h or HAase at 37 °C for 1 h and separated by 4–15% SDS-PAGE. The control of PTX3 and the control of IαI (after NaOH treatment) were mixed just before loading to the same well of the gel. The immunoblot was first probed with rat anti-PTX3 antibody (A) and then stripped and reprobed with mouse anti-HC1 antibody (B). M, protein ladder markers. C, the AM HC-HA complex with or without HAase digestion was separated by 0.5% agarose gel, followed by staining with 0.005% Stains-all dye overnight at 25 °C with light protection. HA was visualized as bluish bands after destaining in water and exposure to room light for 6 h. HMW HA was used as a positive control. M, HA ladder markers.