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. 2016 Jun 9;12(6):e1005696. doi: 10.1371/journal.ppat.1005696

Fig 1. Identification of ppe10::tn as a PE_PGRS supersecretor mutant and its gene product as an ESX-5 substrate.

Fig 1

A, B) Double filter analysis of wild-type M. marinum E11 (wt), the identified transposon mutant ppe10::tn and the complemented ppe10::tn-pSMT3::mmar_0761 (ppe10-C) strain. The double filter of a bacterial plate culture (A), or of single colonies (B) was stained with a monoclonal antibody directed against the PGRS domain. C) PPE10 is secreted in an ESX-5 dependent manner. Immunoblot with protein preparations of wild-type M. marinum E11, the ESX-5 secretion mutant espG 5::tn and the identified ppe10::tn mutant, all containing a plasmid encoding HA-tagged PPE10. The different protein preparations include cell pellets not treated with detergent Genapol X-080 (P -), cell pellets treated with Genapol X-080 (P +), Genapol X-080 supernatant fractions (S +) and culture filtrate fractions (S -). Depicted are the immunoblots stained with anti-HA antibodies or control antibodies (anti-GroEL as cell pellet marker and PE-PGRS as cell surface proteins). Full length PPE10-HA can be observed as a band with an apparent molecular weight of 58 kDa.