The NMDA-R functions as an anti-inflammatory tPA signaling receptor in macrophages. (A) RT-PCR analysis comparing NMDA-R subunit mRNA expression in BMDMs and, as a control, extracts of mouse brain. (B) IF microscopy to detect the NMDA-R NR1 subunit in BMDMs. Nuclei are counterstained with DAPI. Primary antibody was omitted in control studies. (C) BMDMs were pretreated for 30 min with MK801 (1.0 µM), as is indicated, and then with LPS (0.1 µg/mL), h-EI-tPA (12 nM), or both for 1 h, as is indicated. Samples were analyzed by immunoblot analysis to detect phospho-IκBα, total IκBα, and β-actin. (D) BMDMs were pretreated with MK801 (1.0 µM) or vehicle and then with h-EI-tPA (12 nM), RAP (150 nM), or vehicle for 1 h, as is indicated. Immunoblot analysis was performed to detect phospho-IκBα, total IκBα, phospho-ERK1/2, and total ERK1/2. (E-F) BMDMs were preincubated with TLR4-Ab (5 µg/mL) for 3 h or with MK801 (1.0 µM) for 30 min. The cells were then treated with LPS (0.1 µg/mL), h-EI-tPA (12 nM), or both, or with vehicle. RT-qPCR was performed to determine mRNA levels for TNFα (E) and IL-1β (F) (mean ± SEM; n = 4). **P < .01, ***P < .001.