Example sentences for: tlk
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The Taq I fragments of HL60 cDNA were ligated with the CT adapter at 16°C overnight with 3 μl (9 units) of T4 ligase (Promega, Madison, WI) while the Taq I fragments of HL60/TLK286 cDNA were ligated with the TT adapter.
Cellular response to chronic TLK286 exposure included a two-fold decrease in GSTP1-1 protein levels, confirming a GSTP1-1 mediated mechanism of activation [ 7 ] . Mouse embryo fibroblast cells from GSTP1-1 deficient animals elevated the expression of signal-regulated kinases ERK1/ERK2 and reduced the doubling time of cell proliferation [ 8 ] . That paper also found that a clone of HL60 cells resistant to TLK199, an inhibitor of GSTP1-1, has been shown to have elevated activities of c-Jun NH 2 -terminal kinase (JNK) and ERK1/ERK2.
For example, the stress kinase JNK1 was activated during TLK286 induced apoptosis [ 17 ] and JNK is known to be activated during apoptosis induced by a variety of stress stimuli [ 18 ] . However, in HL60/TLK286 resistant cells, we observed an overexpression of two phosphatases PP2 (AA599092) and MKP5 (AA056608) involved in JNK dephosphorylation and inactivation (Table 2).
Briefly, ten μg of total RNA was reverse-transcribed into single stranded cDNA with oligo(dT) 12-18 . Then the double stranded cDNA for HL60 and HL60/TLK286 was generated with the cDNA Synthesis System (Life Technologies, Rockville, MD).
Although the interaction among GSTP1-1 and genes of proliferation and kinase pathways needs to be verified and the roles of other differential genes in the TLK286 resistant cell line need further investigation, the expression pattern generated with ADGE microarray provides a blueprint for designing further experiments.
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