Discrimination of M1 and M2 macrophages via CD38 and Egr2 flow cytometry assay.

<p>A. Experimental design. BMDMs were differentiated for 24 hours in M1 conditions and tagged with CD11b-V450. Another set of BMDM was differentiated in M2 conditions and tagged with CD11b-PE. The M1 and M2 populations were then mixed prior to staining with surface CD38 and intracellular Egr2. CD38⁺Egr2^- (putative M1) and CD38^-Egr2⁺ (putative M2) gates were drawn and the percentage of cells in each gate that originated from M1 (V450 tagged) or M2 (PE tagged) cultures was calculated. B. Representative flow cytometry data of CD38 and Egr2 staining in tagged M1 and M2 populations as indicated in A. C. Quantification of the results from B (n = 3, representative of three independent experiments). D. M0, M1 and M2 cells were stained with either CD11b-V450 or CD11b-PE and subsequently stained with CD38 or Egr2 to confirm that CD11b-V450 or -PE tagging did not alter CD38 or Egr2 staining. ****p<0.0001.