Fig. 1.

PLAUR Is Highly Expressed in Myeloid Cells and Localized at the Cell Membrane. (A and B) Total RNA was extracted from primary monocytes, MDMs, MDDCs, THP-1, PMA-treated THP-1, stimulated or resting CD4⁺ T cells, established cell lines 293T, HeLa, and Jurkat cells. PLAUR transcript levels were measured using quantitative PCR and then normalized to GAPDH levels (A), and data are plotted as mean ± SEM of three independent experiments. Meanwhile, western blotting was conducted to assess the level of PLAUR and GAPDH (B). (C and D). 293T cells (C) and stimulated CD4⁺ T cells (D) exogenously expressed vectors encoding GFP-tagged, non-tagged PLAUR, or GFP. At 48 h after transfection in 293T cells or 96 h after lentiviral vector transduction in CD4⁺ T cells, viable cells were harvested and stained with mouse anti-PLAUR or isotype (IgG) antibodies to measure the levels of the C-terminal GFP-tagged PLAUR and non-tagged PLAUR at the cell surface using a PE-conjugated goat anti-mouse antibody (E). MDMs or MDDCs were lysed, and membrane-bound and cytosolic proteins were separated to assess the levels of PLAUR, Na⁺/K⁺-ATPase, and GAPDH using western blotting with specific antibodies. All micrographs or blotting data are representative of three independent experiments.