Fig. 3

Calcium limiting condition enhances the secretion of Bg_9562 protein into extracellular milieu in a T3SS dependent manner. a Silver stained SDS-PAGE profile of proteins isolated from the culture supernatant of NGJ1 grown in PDB (lane 1), PDB + EGTA (lane 2) and PDB + CaCl₂ (lane 3). While the lane 4, 5 and 6, represents the protein isolated from culture supernatants of T3SS mutant strain of NGJ1 (NGJ12) grown in PDB, PDB + EGTA and PDB + CaCl₂, respectively. b Western blot analysis (using Bg_9562 specific polyclonal antibody) of proteins isolated from the culture supernatant of NGJ1 grown in PDB (lane 1), PDB + EGTA (lane 2), and PDB + CaCl₂ (Lane 3). Lane 4, 5 and 6 represent the protein isolated from the bacterial pellet grown in PDB, PDB + EGTA and PDB + CaCl₂, respectively. Lane 7 contains purified Bg_9562 protein obtained from the recombinant E. coli strain. c Western blot analysis of total protein isolated from the culture supernatant of wild type NGJ1 grown in absence (lane 1) or presence of EGTA (lane 2). While lane 3 and 4 represent proteins from the culture supernatant of NGJ12 grown in absence or presence of EGTA, respectively. Lane 5 contains purified Bg_9562 protein from the recombinant E. coli strain. M reflects pre-stained protein ladder