The Details You Havent Read About PTPRJ

The cells were incubated for 15 min at room temperature in the dark, and 400 ��l of binding buffer was added. The processed ells were analyzed with FACScan (Becton-Dickinson, USA). The PC-3 cells were gated separately according to their granularity and size on forward scatter (FSC) versus Side Scatter (SSC) plots. Early and late apoptosis was evaluated on fluorescence 2 (FL2 for propidium iodide) versus PTPRJ fluorescence 1 (FL1 for Annexin) plots. Cells stained with only annexin V were defined as being in early apoptosis; cells stained with both annexin V and propidium iodide were defined as being in late apoptosis or in a necrotic stage. Cells were analyzed with a flow cytometer (FACSCalibur, BD, Vienna, Austria). To investigate the expression level of multidrug-resistance genes Afatinib in vivo in transplanted tumor tissues, we employed a reverse transcriptase-polymerase chain reaction (RT-PCR) assay using mdr1, BCR/ABL and sorcin primers. Total RNA was extracted from the frozen tumor tissues. Integrity of RNA was demonstrated by a high-resolution gel method. After the reverse transcription, mdr1, BCR/ABL, sorcin and GAPDH primers were used for cDNA amplification. PCR products were electrophoresed on agarose gels containing ethidium bromide and visualized by UV photography. Primer sequences for mdr1, BCR/ABL, sorcin and GAPDH are shown in Table 2. To assess the role of mdr1, sorcin, BCR/ABL genes in drug resistant K562/ADM tumor cells, mdr1, sorcin, BCR/ABL gene expression was also evaluated in non-drug resistant K562 cells in the same manner as in K562/ADM tumor cells. Cells of the control group and three HSS groups were lysed by boiling in 2��SDS sample buffer. Lysate proteins (50 ��g) were separated by SDS-PAGE and transferred to nitrocellulose. Western blots were serially probed with combinations of antibodies to mdr1, sorcin, BCR/ABL and caspase-3 (Santa Cruz Biotechnology, Santa Cruz, CA), ?-catenin (to control for loading). Each experiment was repeated several times with different batches of proteins, and representative blots are shown. Data are expressed as means �� SD. Dasatinib datasheet SAS6.12 software was used for data analysis. Numerical data were analyzed with one-way ANOVA. Categorical data were analyzed with Chi-square test. P