Skip to main content
. 2020 Mar 17;295(18):5960–5969. doi: 10.1074/jbc.RA119.011682

Figure 2.

Figure 2.

Homologously expressed EsxA_1/EsxB_1 require co-expression and secretion of PE35/PPE68_1 for efficient secretion via the ESX-1 system. A, a schematic representation of the different constructs used are shown on the left. The four genes encoding the M. marinum ESX-1 substrates PE35/PPE68_1 (in blue) and EsxB_1/EsxA_1 (in pink) are either expressed from separate hsp60 promoters (I and II) or co-expressed under the same hsp60 promoter (III). Immunoblot analysis was performed of the cell pellet and culture supernatant fractions of WT M. marinum using an HA antibody to detect EsxA_1–HA, a FLAG-antibody to detect PPE68_1–FLAG, an EsxA antibody, detecting both endogenous and exogenous EsxA paralogs, and a GroEL2 antibody to detect the intracellular control protein GroEL2. B, a schematic representation of the different constructs used is shown on the left; deletions are indicated by ×. Immunoblot analysis using the same antibodies as under A to analyze secretion by WT and the ESX-1 mutant. In all blots, equivalent OD units were loaded; 0.2 OD for pellet and 0.5 OD for supernatant fractions. Numbers indicate two independent M. marinum colonies carrying the same construct. E, empty strain.