im inducing human microglial cell lines with LPS what i am wondering is that whether the activation of NFKb by LPS is dose-dependent or time dependent? because in latest test i made i had induced the activation of p-p65 in course of time 10,30,60 min with 1 ug/ml of LPS however the result using western blot is that increased NFkB expression in 10 and 30 min, but on the 60 min the expression is closed with basal state. so my quation is that whether the expression of NFkB will recover(high expressed of p-p65) on 24hr induction of LPS using same dose?
and in the same test i find interesting is that there are no difference in expression in IKb-alpha with LPS treatment and basal state.
I am not sure in your cell lines, but in BMDM it is both dose and time dependent. However, it is very easy to overstimulate them and cause sufferance and death , over-stimulation and cell death are also dose and time dependent, it is usually possible to appreciate the change of morphology at the microscope
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