For immunostaining, samples were stained with Alexa Fluor secondary antibodies.
After washing with PBS, samples were stained with secondary antibodies as above.
For exclusion of death cells, samples were stained with Hoechst 33342 (Invitrogen).
At each point samples were stained with PI, and incorporation was quantified by flow cytometry.
To exclude dead cells, the samples were stained with propidium iodide (BD Bioscience, Heidelberg, Germany).
The samples were stained with three different MitoTracker dyes, similar to Hbt. salinarum, and were examined using a confocal fluorescence microscope.
Samples were stained with SYBR Green I (Molecular Probes) and yellow-green latex beads (Polyscience, 0.5 μm) were used as an internal standard.
The samples were stained with Hoechst 33342 (Invitrogen, diluted to 1 μg/ml), mounted in ProLong Gold mounting medium (Life technologies P36930).
At the indicated time points, samples were stained with Calcofluor and the numbers of fluorescent cysts and unstained amoebas were counted.
The samples were stained with calcofluor white.
Finally, deparaffinized, the samples were stained with H&E (hematoxylin-eosin) and microscopically analysed.
