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TO-PRO-3 iodide (Invitrogen, http://www.invitrogen.como) was used as a marker for nucleus.
Tissues were incubated with primary antibodies overnight at 4°C, followed by fluorochrome-conjugated secondary antibodies for 60 min at room temperature, and then counterstained with marker for nucleus 4′, 6-Diamidino-2-phenylindole dihydrochloride (DAPI) and/or FluoroMyelin.
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XFM analysis of mouse mammary gland samples demonstrated elemental distributions for sulfur (correlated to overall cell thickness) and phosphorus (utilized as a marker for nuclei) to be typical of eukaryotic cells [27].
Results of ASMA were verified by immunofluorescence in colocalization with DAPI as a marker for nuclei.
Using DAPI (blue) and MitoTracker (red) as marker for the nucleus and mitochondria respectively, we find that GST-CG4567 witheuTPthexclusivelyivelocalizeszes to the nucleus (Figure 6J).
Colocalization is visible as yellow in the merged image, with 4',6-diamidino-2-phenylindole (DAPI, blue) as a general marker for the nucleus.
The former provides a marker for all nuclei, thus facilitating all cells to be followed at successive stages; the latter provides a marker to follow the pluripotent cells before the egg cylinder stage and germ cells thereafter (Supplementary Fig. S2a,b).
The dramatic nearly complete elimination of pyramidal CA neurons was confirmed by Nissl staining (data not shown) and by IHC for NeuN as marker for neuronal nuclei (Figure 3A, lower panels).
The cellular location of SPION was stained with the plasma membrane marker CellMask (Invitrogen), lysosome marker LysoTracker Green (Invitrogen) and a marker for cell nuclei, Hoechst 34580 (Invitrogen).
We combined these stains with a fluorescent marker for cell nuclei (SYTOX Green), and an antibody directed against α-smooth muscle actin followed by a fluorescently conjugated secondary antibody to identify smooth muscle cells.
Immunostaining for a variety of neuronal markers, including pan-neuronal markers (Map2, TuJ1), interneuron subtype markers (calbindin and calrectinin), a marker for proliferating nuclei (phosphoHistone3), and for glial and radial glial markers (GFAP and RC2) all showed no significant differences between mutant and control littermate brains (data not shown).
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