Figure 4
Localization of PnxIIIA and the protein interaction analysis of rPnxIIIA. (A) Western blotting analysis of the cell fraction prepared from P. pneumotropica ATCC 35149 cells and culture by using anti-rPnxIIIA IgG. Lanes: 1, SC fraction; 2, IM fraction; 3, OM fraction; 4, UC fraction. (B) Western blotting analysis of the cell fraction prepared from P. pneumotropica ATCC 35149 cells and culture by using anti-rPnxIIIE IgG. Lanes: 1, SC fraction; 2, IM fraction; 3, OM fraction; 4, UC fraction. (C) Western blotting analysis of rPnxIIIA by using anti-rPnxIIIA IgG after cross-linking with only rPnxIIIA or the rPnxIIIE protein and IP with anti-rPnxIIIA IgG. Lanes: 1, rPnxIIIA without cross-linking; 2, 20 μg of rPnxIIIA alone cross-linked with 50 mM BS3 for 60 min and immunoprecipitated; 3, mixture of both rPnxIIIA and rPnxIIIE proteins without cross-linking; 4, 20 μg of both rPnxIIIA and rPnxIIIE proteins cross-linked with 50 mM BS3 for 60 min and immunoprecipitated. (D) Western blotting analysis of rPnxIIIA by using anti-rPnxIIIA IgG after different treatment times with rPnxIIIA alone cross-linked with 50 mM BS3 and immunoprecipitated with anti-rPnxIIIA IgG. Lanes: 1, rPnxIIIA without cross-linking; 2, rPnxIIIA with cross-linking for 5 min; 3, rPnxIIIA with cross-linking for 10 min; 4, rPnxIIIA with cross-linking for 30 min; 5, rPnxIIIA with cross-linking for 60 min. Arrows indicate the position of the bands that appeared.