Fig 6. Fluorescence confocal microscopy images of mouse primary cardio fibroblasts infected with T. cruzi show the return of Tc24 expression.

Two images were acquired that represent different stages of infection. In A1-A5 two infected cardiac fibroblasts can be seen with only approximately 20 and 6 amastigotes inside, representing stage IV: amastigotes with several (2–4) divisions. In B1-B5 an infected cardiac fibroblast can be seen hosting 60 amastigotes inside, which represents stage V: transformation from amastigote back to trypomastigote. 1) In blue, DAPI nuclear stain. 2) In magenta, detection of the amastigote specific protein SSP-4 using a specific IgG1 antibody followed by staining with anti-IgG1 Alexa Fluor 488. 3) In yellow, detection of Tc24 using biotinylated Tc24-C4/884 followed by straining with streptavidin–Alexa Fluor 555. 4) In cyan, filamentous actin stained with Phalloidin iFluor 647. 5) Merged image of 1–4. Pixel size of images: 120 nm x 120 nm.