Research Paper Volume 16, Issue 19 pp 12928—12951

Figure 5. PACSIN1 mediates the effects of GATA2 downregulation on macrophage by regulating phagolysosome formation. RAW264.7 cells were co-transfected with or without siGata2 and Pacsin1 overexpression plasmids for 48 h. mRNA levels of Gata2, Pacsin1 and Synj1 were assessed by qPCR (A) and protein levels of GATA2, ABCA1, PACSIN1, SYNJ1, EEA1, and LAMP1 were assessed by western blotting (B) and immunofluorescence staining (C) in RAW264.7 cells from each group. mRNA levels of Cd163, Cd209, Marco, Stab2, Sirpa, Lamp2 and Gpnmb were measured by qPCR (D) in RAW264.7 cells from each group (siNC, black bars; siGata2-1, gray bars; siGata2-2, white bars; siGata2-1+Pacsin1-OE, teal bars; siGata2-1+Pacsin1-OE, asparagus bars). Phagocytosis assessment was done by Cell Meter^™ Fluorimetric Phagocytosis Assay Kit (E). Scale bars = 50 μm. ^*P < 0.05 or ^**P < 0.01 vs. siNC and ^#P < 0.05 or ^##P < 0.01 vs. siGata2.