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Figure 2 | Journal of Translational Medicine

Figure 2

From: Epigenetic inactivation of mir-34b/c in addition to mir-34a and DAPK1 in chronic lymphocytic leukemia

Figure 2

Methylation of miR-34b/c , miR-34a and DAPK1 in CLL cell lines. (A) In CLL cell lines, MEC1, 232B4, I83-E95 and WAC3CD5+ were completely methylated and MEC2, HG3 and CLL-AAT were completely unmethylated for miR-34b/c. I83-E95 was completely methylated, MEC1 was partially methylated, but MEC2, 232B4, CLL-AAT, HG3 and WAC3CD5+ were unmethylated for miR-34a. Moreover, MEC1 was completely methylated, and other six cell lines were unmethylated for DAPK1. (B) Stem-loop qRT-PCR analysis of the mature miR-34b expression in 7 CLL cell lines. ∆Ct, Ct miR-34b-Ct RNU48. (C) In CLL cell lines, MEC1 cells showed no DAPK1 mRNA expression while other six cell lines had detectable DAPK1 mRNA levels. (D) Sequencing analysis of the semi-quantitative RT-PCR products for the detection of DAPK1. MW: Marker; B: Reagent blank; N1: Normal donors; NoRT: Negative control without reverse transcriptase; PC: positive control with methylated DNA. (E) Western blot analysis of DAPK1 in MEC1, MEC2, 232B4, CLL-AAT and WAC3CD5+ cells. WAC3CD5+ cells were used as positive control. Anti-actin protein was regarded as the endogenous normalizer and the relative DAPK1 protein level was shown in the bottom row.

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