Research Paper Volume 16, Issue 19 pp 12928—12951

Figure 6. GATA2 improves macrophage inflammatory phenotype and phagocytosis against CSE through transcriptional activation of ABCA1 and PACSIN1. RAW264.7 cells were pre-incubated with or without CSE. After 24 h, they were transfected with or without Gata2 overexpression plasmids for another 48 h. mRNA levels of Abca1, Tlr4, Myd88, Megf10, Gulp1, Pacsin1, and Synj1 were assessed by qPCR and their protein levels, GATA2 and Flotillin1 were added, which were measured by western blotting in RAW264.7 cells from each group (A). mRNA levels of Cd80, Cxcl1, Ccl2, Il23, Cxcl10, Cxcl11, Cxcl12, Mmp9, Mmp12 and Tgfb1 were assessed by qPCR (B) in RAW264.7 cells from each group (NC, black bars; CSE, gray bars; CSE+Gata2-OE, white bars). Phagocytosis assessment was done by Cell Meter^™ Fluorimetric Phagocytosis Assay Kit (C). Scale bars = 50 μm. ^*P < 0.05 or ^**P < 0.01 vs. NC and ^#P < 0.05 or ^##P < 0.01 vs. CSE. Schematic drawing shows loss of GATA2 in macrophage during COPD is associated with the loss of ABCA1 and PACSIN1. GATA2 directly regulates ABCA1, which is essential for pro-inflammation via TLR4/MYD88 and for phagocytosis via MEGF10 and GULP1. Together with SYNJ1, the decline in PANCIN1 expression, which is another target gene of GATA2, directly hindered the formation and maturation of phagolysosome in macrophages (D).