Figure 2

Anti-inflammatory molecules IL-10, Arg1, Il13ra2, and Mrc1 are highly expressed in REPELL-microglia. (a–c) C8-B4 microglia were treated with low-dose LPS (1 ng/mL) one or three times (n = 3, in triplicate), and relative mRNA expression of anti-inflammatory molecules was measured by real-time RT-PCR using the 2^−∆∆Ct method 4 h after the last LPS treatment. Data were normalized to GAPDH and expressed as the relative fold change over untreated cells. (a) Cytokines and metabolic enzymes, (b) cell surface receptors, and (c) transcriptional regulators. (d) Secretion of IL-10 was promoted by repetitive low-dose LPS in microglia. The levels of IL-10 in culture supernatant were measured by ELISA 30 h after the last LPS treatment. (e) Mrc1 expression on cell surfaces was promoted by repetitive low-dose LPS in microglia. The cell surface antigen Mrc1 was stained, and the mean fluorescence intensity (MFI) of 50,000 counted cells was assessed 6 h after the last LPS treatment using a Beckman Coulter Gallios flow cytometer and Kaluza software. Data are expressed as the relative fold change over untreated cells. Mean ± SE of each group are shown. Data are representative of three independent experiments. LPS x0, no treatment; LPS x1, single treatment with low-dose LPS; LPS x3, treatment with low-dose LPS three times every 24 h. *p < 0.05 for one-way ANOVA with Tukey’s post-hoc correction for multiple comparisons. n.s., not significant.