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Imaging was performed using a Keyence fluorescence microscope (BZ-X710, Keyence Corporation of America, Itasca, IL) with a 20 objective in both DAPI and RFP channels

Quantitative determination of Bcl-2 in FACS-sorted single cells by laser scanning cytometry (LSC): Comparison with flow cytometric and Western blot analysis

The GST cDNA sequence from pGEX-6P-2 (GE Healthcare) was PCR amplified with primers containing NcoI and KpnI restriction sites at the 5' and 3' extremities, respectively

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Image-Pro Plus 6.0 software was used to measure the area of IHC DAB staining to obtain the integrated optical density (iod) and staining area (area) values