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Cell attachment and proliferation was further confirmed by nuclear staining using 4′,6-diamidino-2-phenylindole (DAPI).
Primary and secondary antibody incubations were subsequently undertaken at 37°C for 1 hr each followed by nuclear staining using DAPI (1 µg/ml) diluted 1/500 in water.
Apoptosis of podocyte was detected by nuclear staining using DAPI.
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One could use more specific stains than the nuclear stains used here.
Purity was confirmed by nuclear staining with DAPI using fluorescence microscopy.
After morphological assessment by nuclear staining, extent of apoptosis was quantified using the TUNEL assay (described below).
Apoptosis was measured by nuclear staining and Annexin V-PI staining.
Nuclear translocations were confirmed by nuclear staining with Hoechst H33258.
Cell density was determined by nuclear staining with crystal violet.
For PP1γ, the expression was evaluated by nuclear staining.
For HIF-1 α analysis, positive expression was defined by nuclear staining and scored as follows: 0=absence of staining, 1=<25% staining, 2=25 50% staining, 3=50 75% staining, 4=>75% staining.
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