Figure 1

Pro-inflammatory molecules Nos2, Ccl1, IL-12B, and CD86 are highly expressed in REPELL-microglia. (a) The experimental schedule of LPS treatment. C8-B4 microglia cells were treated with low-dose LPS (1 ng/mL) one or three times (n = 3, in triplicate). (b,c) relative mRNA expression of pro-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 a relative fold change over untreated cells. (b) Cytokines and metabolic enzymes, (c) cell surface receptors, and (d) transcriptional regulators. (e) The secretion of IL-12B was promoted by repetitive low-dose LPS in microglia. Cytokine levels of TNF-α, IL-12B, and IL-6 in culture supernatant were measured by ELISA 30 h after the last LPS treatment. (f) CD86 expression on cell surfaces was promoted by repetitive low-dose LPS in microglia. The cell surface antigen CD86 was stained, and the 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.