I have been trying to block KDM6B and KDM5B with to chemical blockers (subretinal injection-rat). I could not detect any changes in treated eye vs control eye with Western Blotting for H3k27me3 and H3k4me3.
Can anyone help me with this issue?
Thank you for help.
Raquel Silva
Are the demethylase blockers specific to those enzymes? If yes, there are multiple members of both KDM5 and KDM6 families, could it be they are compensating? Did you also tried different doses ? You might want to try to scale up?And how long after injection did you collected the tissue? You could try to optimise your blockers in a cell line that is dividing, and you could also try to use CRISPR-i to inactive the enzymes.
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