I have been working with NLRP3 inflammasome signalling and have been using RAW 264.7 murine macrophage cells stimulated with LPS to do so. I have recently come across several sources asserting that RAW 264.7 cells do not express ASC protein and thus are incapable of NLRP3 activation. However, I have also found sources that use RAW 264.7 for NLRP3 studies. Further complicating matters, some data from my lab has supported NLRP3 activation in RAW cells; but more recently, I have been unable to find any trace of typical NLRP3 response. What is going on? Do RAW 264.7 cells express ASC or not? Any expertise or help would be much appreciated.
RAW 264.7 cells DO NOT express ASC. In fact, for Cell Signaling’s murine anti-ASC antibody it is a negative control (https://www.cellsignal.com/products/primary-antibodies/asc-tms1-d2w8u-rabbit-mab-mouse-specific/67824). I have seen some people transfect an ASC construct into RAW 264.7 cells to study the inflammasome with them however (https://journal-inflammation.biomedcentral.com/articles/10.1186/1476-9255-7-23). If you wanted to simply study how transcriptional priming is affected by certain conditions then RAW 264.7 cells are fine, but, if you’re interested in the activation of the inflammasome, then I would suggest using another cell line (e. g., J774.2 or THP-1 cells) or primary cells (Bone marrow-derived macrophages or peritoneal macrophages). I hope this helps; good luck!
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