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Fig. 3 | Journal of Neuroinflammation

Fig. 3

From: Human microglia and astrocytes constitutively express the neurokinin-1 receptor and functionally respond to substance P

Fig. 3

U87-MG astroglioma cells constitutively express NK-1R, and expression of this receptor is increased following exposure to bacterial components. Panel a U87-MG cells were untreated (C) for 24 h or exposed to LPS (5 ng/mL) for 2, 12, and 24 h, and the level of expression of mRNA encoding NK-1R was determined by RT-PCR (n = 4). Panel b Cells were untreated (0) or exposed to LPS (5 or 10 ng/mL) for 24 h and protein expression of fNK-1R, tNK-1R, and the housekeeping gene product β-actin, was determined by immunoblot analysis (n = 3). Expression in mouse heart tissue (mh) is included as a positive control for tNK-1R. Panel c Cells were untreated for 24 h (C) or exposed to LPS (5 ng/mL) for 2, 12, and 24 h, and the level of fNK-1R protein expression was determined by immunoblot analysis. Data is shown as protein levels, normalized to β-actin expression, relative to NK-1R expression in untreated cells (n = 4). Panel d Cells were untreated (0) or exposed to LPS (5 ng/mL) or bacterial flagellin plus PAM3Cys at 100 ng/mL and 500 ng/mL (lo) or 200 ng/mL and 1000 ng/mL (hi), respectively, and cell surface NK-1R expression was determined at 2 h by flow cytometry. Data is shown relative to cell surface NK-1R expression on untreated cells (n = 5). Data is expressed as the mean ± SEM and asterisks indicate statistically significant differences between untreated and treated cells (p < 0.05)

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