Fig. 3
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- ZDB-FIG-230430-85
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- Saelens et al., 2022 - An ancestral mycobacterial effector promotes dissemination of infection
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Dissemination in vivo is dependent on esxM
(A) A macrophage infected with WT M. marinum departs from an early granuloma and migrates above the midline (dashed white line). Scale bar, 50 μm. (B) Representative images from WT M. marinum, ΔesxM, and ΔesxM:esxM infections in larval zebrafish. 2 dpf zebrafish with red fluorescent macrophages are infected ventrally in the caudal vein with cerulean-expressing M. marinum. Scale bar, 500 μm. (C) At 5 dpi, ΔesxM demonstrates a reduced frequency in dissemination above the midline (white dashed line) to the dorsal side of the animal adjusting for burden across all three groups. One-way ANOVA with Tukey’s post-test, data from three biological replicates, mean ± SD shown. n > 35/experiment. (D) Co-infection with WT and ΔesxM M. marinum results in larval granulomas populated by both strains. Time-lapse imaging reveals WT M. marinum egressing from the granuloma (tracking lines), while ΔesxM remains confined within. White arrows indicate direction of egress. Scale bar, 50 μm. (E) Projection of time-lapse imaging in (D) showing complete range of movement for macrophages infected with each strain over 220 min; Scale bar 50 μm. (F) Quantification of rate of egress for WT and ΔesxM M. marinum from co-populated granulomas. Long-term monitoring of 4 mixed granulomas from 4 independent animals. Student’s paired t test, mean ± SD shown. (G) Migration of RAW 264.7 cells expressing fluorescent cerulean control or EsxM through transwell membranes in the presence and absence of chemokine. One-way ANOVA with Bonferonni’s post-test. *p < 0.05, **p < 0.01, ***p < 0.001. See also Figure S3, Table S4, and Videos S1 and S2. |
Reprinted from Cell, 185(24), Saelens, J.W., Sweeney, M.I., Viswanathan, G., Xet-Mull, A.M., Jurcic Smith, K.L., Sisk, D.M., Hu, D.D., Cronin, R.M., Hughes, E.J., Brewer, W.J., Coers, J., Champion, M.M., Champion, P.A., Lowe, C.B., Smith, C.M., Lee, S., Stout, J.E., Tobin, D.M., An ancestral mycobacterial effector promotes dissemination of infection, 4507-4525.e18, Copyright (2022) with permission from Elsevier. Full text @ Cell