FIG. 7.
Western blot analysis of invB-dependent expression and secretion of SopE2M45 in SL1344 derivative strains and of SopE2 in ATCC 14028s derivatives. (A) SL1344(pM256), SB566(pM256), M568(pM256), and M568(pM256, pM249) were grown under SPI-1-inducing conditions (see Materials and Methods). One hundred microliters of a whole culture, pelleted bacteria from 100 μl of a culture, and proteins recovered from 1 ml of a culture supernatant were analyzed by Western blotting by using an anti-M45 (α-M45) antibody. α-OmpC, anti-OmpC antibody. (B) SL1344(pM226), SB161(pM226), and M568(pM226) were grown under SPI-1-inducing conditions (see Materials and Methods). One hundred microliters of a whole culture and proteins recovered from 500 μl of a culture supernatant were separated by SDS-PAGE and analyzed by Western blotting by using an anti-M45 (α-M45) antibody. α-OmpC, anti-OmpC antibody. (C) CS401, M622, PB502, and PB502(pM250) were grown under SPI-1-inducing conditions. Pelleted bacteria from 200 μl of a culture and proteins recovered from 1 ml of a culture supernatant were analyzed by Western blotting by using anti-SopE2 (α-SopE2) and anti-SipC (α-SipC) antisera. The blots were reprobed by using an anti-OmpC (α-OmpC) antibody to control the amount of protein loaded.