ppat.1009410.s002.tif (4.06 MB)

Differential intracellular trafficking between Mtb and Mbv.

figure

posted on 2021-03-15, 17:39 authored by Christophe J. Queval, Antony Fearns, Laure Botella, Alicia Smyth, Laura Schnettger, Morgane Mitermite, Esen Wooff, Bernardo Villarreal-Ramos, Waldo Garcia-Jimenez, Tiaan Heunis, Matthias Trost, Dirk Werling, Francisco J. Salguero, Stephen V. Gordon, Maximiliano G. Gutierrez

hMϕ or bMϕ infected with Mtb-RFP or Mbv-RFP (A, B, E and F) or Mtb-GFP or Mbv-GFP (C and D) for 24 h. Samples were fixed and fluorescently stained for the late endosomal marker Lamp-1 (A and B), for the pH sensitive dye LysoTracker DN99 Red (LTR) (C and D) and for the autophagic marker LC3B (E and F). For each fluorescent confocal image, the cell nuclei were stained with DAPI. Positive association of bacteria with the different markers, delimited by a white square, are magnified and displayed at the top right corner and the right-hand side of each image. Scale bars represent 10 μm. Graphs represent the quantification of the marker association with Mtb or Mbv ± SEM from three independent experiments. Each dot represents the mean relative fluorescent intensity of the cellular marker with a single or distinct bacteria group. The population within each dotted red box corresponds to the percentage (± STD) of bacteria positive for the marker tested.

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