I am trying to induce necroptosis in macrophages differentiated from Hoxb8 cells. I have attempted this by incubating the cells with the z-VAD pan caspase inhibitor (10uM) and TNF-alpha (10ng/ml) for 4h, but to no success. I am assessing the level of necroptosis using the Sytox Green nuclear stain on PFA-fixed cells and imaging with a microscope.
Thank you