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. Author manuscript; available in PMC: 2018 Mar 19.
Published in final edited form as: Methods. 2016 Sep 15;112:91–104. doi: 10.1016/j.ymeth.2016.09.007

Figure 5. Detection of H37Rv-mCherry MTb in early and/or late phagosomal compartments of infected human THP-1 cells.

Figure 5

Wild type THP-1 cell were infected with H37Rv-mCherry MTb for 24 hours, fixed in 4% PFA, and stained with AB to Rab5 or Rab7 or Rab5 and CD107a and acquired on IS-X Mark II, as described in Ranjbar et al [90].

5A: Representative images of internalized MTb-mCherry in wild type THP-1 cells 24 hr after infection. Cells were labeled with Rab5 antibody and stained with DAPI as above. Images shown from left to right: BF image (Grey), DAPI (Purple), MTb-mCherry fluorescence (Red), Rab5 (Green), and a merged image showing colocalisation of MTb-mCherry fluorescence and Rab5. BF similarity coefficient values are shown in yellow font.

5B: Representative images of internalized MTb-mChery in wild type THP-1 cells that were labeled with Rab7 and stained with DAPI as above. From left to right: BF image (Grey), DAPI (Purple), MTb-mCherry fluorescence (Red), Rab 7 (Blue), and a merged image of MTb-mCherry fluorescence and Rab7 with BF similarity coefficient shown in yellow font.

5C: Representative images of internalized MTb-mCherry in wild type THP-1 cells infected with MTb-mCherry and labeled with Rab5 and CD107a (LAMP1) as above. From left to right: BF image (Grey), MTb-mCherry fluorescence (Red), CD107a (Dark blue), Rab5 (Green), and a merged image of MTb-mCherry fluorescence and CD107a with BF similarity coefficient in yellow font, a merged image of MTb-mCherry fluorescence and Rab5, and a merged image of MTb-mCherry fluorescence, CD107a and Rab5.