Gene expression of methylated gene
Posted 20 April 2010 - 10:35 AM
I am new to DNA methylation studies and I would appreciate a little help on this. I'm just trying to standardize my techniques for RT-PCR and at the moment, I am looking at gene expression of a known methylated gene (GSTP1) in prostate cells (LNCaP).
What I'm finding a bit strange is that I am seeing expression of the control (untreated) cells in RT-PCR and Western analysis. Could anyone suggest a possible reason for this. There are papers that suggest GSTP1 should be silenced in this cell line. I have also done treatments with 5aza-2'deoxycytidine and see very little change from the control.
Posted 20 April 2010 - 03:32 PM
Posted 20 April 2010 - 11:18 PM
Hope this helps.
Posted 25 April 2010 - 05:27 PM
I do see a slight increase in GSTP1 and RASSF1 expression after 5azaDcd treatment but I still see some expression in the controls. Is this normal? Is there expected to be some expression at the basal level?
Also, is there any type of marker to make sure this is LNCaP cells? I received these cells from someone else so there could be a mistake in its identity.
Posted 25 April 2010 - 10:18 PM