Did you control the level of confluence of your cells, and the quality in terms of nutriments of your medium.

Is your medium (well) buffered?

Is the temperature in your incubator very stable?

What type of cells are you using?

Best regards

Robert

I agree with Robert Kiss. Some minor alterations in cell confluence, pH, nutrient source and quantity, and temperature can lead to alterations of ER stress gene expression. This can also happen with some contaminations that activate cells (mycoplasma and LPS).

Thanks for your reply .

Confluence of the cell i do check.

I am using primary cultured mice glial cells..

media i am using GIBCO hi glucose DMEM with 10% serum..and i never observed change in temperature in my incubator.

Jit

I don't have any experience with glial cells but could it be that your isolation process is activating ER response? I believe your protocol involve dislodging the tissue mechanically and using enzymes to digest tissue. Can you take small samples for each step (doesn't matter cells aren't purified yet) just to check the expression of ER stress genes? Maybe your cell isolation protocol needs adjustment.

I fully sare Andre's remark / suggestion.

Robert