Comparative analysis of RT-qPCR, flow cytometry, and Di-4-ANEPPDHQ fluorescence for distinguishing macrophages phenotypes.

ABSTRACT: This study evaluates the effectiveness of fluorescence microscopy using Di-4-ANEPPDHQ in differentiating macrophage phenotypes (M0, M1, and M2) compared to RT-qPCR and flow cytometry. Using THP-1 monocyte-derived macrophages, we assessed cytokine expression (IL-1β, IL-6, IL-10) via RT-qPCR, surface markers (CD86, CD64, CD206) through flow cytometry, and membrane properties with Di-4-ANEPPDHQ fluorescence. RT-qPCR showed significant differences in cytokine expression: M1 macrophages had elevated IL-1β (p < 0.0001) and IL-6 (p < 0.0001), while M2 macrophages exhibited higher IL-10 levels (p = 0.0030). Flow cytometry revealed distinct surface marker profiles, with M1 expressing high CD64 and M2 showing increased CD206. Di-4-ANEPPDHQ fluorescence indicated membrane order differences: M1 macrop

SUBMITTER: Siniyeh AA 

PROVIDER: S-EPMC12418842 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature