Figure 1
Analysis of the SHP1 gene expression and methylation in various hematopoietic cell lines. (A) Western blot analysis of SHP1 protein expression. Cell lysates extracted from 7.5 × 104 cells cultured under standard conditions were subjected to SDS-PAGE, blotted onto membranes and probed with anti-SHP1 or anti-β actin antibodies. (B) RT-PCR analysis of the SHP1 gene. SHP1 and β actin cDNA were amplified by RT-PCR, then separated by 1.5% agarose gel electrophoresis. N.C. indicates the negative control, which did not contain cDNA. (C) MSP and unMSP assays of the SHP1 promoter in hematopoietic cell lines and normal peripheral blood mononuclear cells (PBMCs). M and U indicate methylation and unmethylation specific PCR, respectively. (D) Restriction enzymes-mediated PCR of the SHP1 promoter region in hematopoietic cell lines and normal PBMC. DNA was digested with HpaII or MspI for PCR.
