KefB inhibits phagosomal acidification but its role is unrelated to M. tuberculosis survival
in host
Garima Khare, P. Vineel Reddy, Pragya Sidhwani and Anil K. Tyagi*
Supplementary Information for Figure 1
Given below are the original gel pictures for the confirmation of the disruption of kefB in M.
tuberculosis. The portions marked as Fig 1B, 1C, 1D and 1E have been used separately in the
final Figure 1.
Legend to Supplementary Figure S1
Figure S1. Localization of M. tuberculosis, MtbΔkefB, MtbΔkefBComp in early endosomes
by employing Rab5 antibodies. RAW 264.7 macrophages were infected with FITC labeled M.
tuberculosis, MtbΔkefB, MtbΔkefBComp and latex beads (green) separately. The cells were then
subjected to staining with Rab5 primary antibodies and counterstained with Texas Red labeled
secondary antibodies followed by observation under confocal microscope. Representative
fluorescent images depict that disruption of kefB leads to a poor accumulation of MtbΔkefB in
early endosomes while the M. tuberculosis and MtbΔkefBComp were found to accumulate in
Rab5 rich organelles (overlap of green and red images appears yellow as shown by arrows).
Latex beads employed as control display poor colocalization with the Rab5 rich early
endosomes. The scale bars depict 5 µm.