Burkholderia cenocepacia BC2L-C Is a Super Lectin with Dual Specificity and Proinflammatory Activity
Figure 8
Localisation of B. cenocepacia soluble lectins in bacterial cells.
A. BC2L-A, -B and –C are secreted into the growth medium. Analysis of concentrated culture supernatants and total cell lysates recovered from B. cenocepacia K56-2 containing the control plasmid pEL-1 (lanes 1 and 5), K56-2 pBC2L-A-FLAG (lanes 2 and 6), K56-2 pBC2L-B-FLAG (lanes 3 and 7) and K56-2 pBC2L-C-FLAG (lanes 4 and 8). B. BC2L-B and –C are released from the bacterial surface upon mannose treatment. Analysis of concentrated supernatants from cells treated with PBS or 50 mM d-mannose and total cell lysates recovered from B. cenocepacia K56-2 pBC2L-A-FLAG (lanes 1), K56-2 pBC2L-B-FLAG (lanes 2) and K56-2 pBC2L-C-FLAG (lanes 3). Western blots were performed using anti-FLAG and anti-RNAP alpha subunit antibodies (cell lysis control). A degradation product from BC2L-C can be detected in the total cell lysates. The arrow highlights the position of the RNAP alpha subunit in total cell extracts. Samples were boiled 10 min prior to loading on 18% SDS-PAGE gels.