Fig. 2: Analysis of secretion proteins in M. abscessus lacking ESX-3 and/or ESX-4. | Communications Biology

Fig. 2: Analysis of secretion proteins in M. abscessus lacking ESX-3 and/or ESX-4.

From: Deletion of ESX-3 and ESX-4 secretion systems in Mycobacterium abscessus results in highly impaired pathogenicity

Fig. 2

Immunoblot analysis of bacterial total lysates (Pellet) and secreted proteins (Supernatant) in Mab S (lane 1), ΔeccC3 (lane 2), ΔeccC4 (lane 3), and ΔeccC3/ΔeccC4 (lane 4). Immunoblots illustrating the secretion profile of EsxH (a), PPE4 (b), EsxT (c), EsxH2 (d), MAB_0664 (e), and MAB_0047 (f). Anti-Ag85 antibodies were used as loading controls to verify that the secretion of Ag85 is not affected by the disruption of ESX-3 or ESX-4. GroEL2 serves as a loading control for total lysates and to validate the absence of bacterial lysis. This figure presents data from experimental replicate 1 only. The secreted proteins were tagged with HA tag and expressed from integrative plasmids. Specific proteins were identified using anti-HA antibodies. The design of the fusion proteins and their tags is shown above each blot: (a) EsxG/EsxH-HA, (b) PE5-PPE4-HA-EspG3-STREP, (c) EsxU/EsxT-HA, (d) EsxG2/EsxH2-HA, (e) MAB_0664-HA, (f) MAB_0046-MAB_0047-HA. g Quantification of the bands present in the supernatant fractions shown in panels (a–f) using the Fiji software (mean gray values) (n = 3). h Secretion analysis of the same substrates in complemented strains expressing either EccC3-mNeonGreen or EccC4-mNeonGreen. Data are mean ± SD.

Back to article page